CN101253404A

CN101253404A - Manipulation and/or detection of biological samples and other objects

Info

Publication number: CN101253404A
Application number: CNA2005800190274A
Authority: CN
Inventors: 咸燉熹; 罗伯特·韦斯特维尔特; 托马斯·亨特; 刘勇; 李学虎
Original assignee: Harvard College
Current assignee: Harvard College
Priority date: 2004-04-13
Filing date: 2005-04-13
Publication date: 2008-08-27

Abstract

Methods and apparatus for manipulation, detection, imaging, characterization, sorting and/or assembly of biological or other materials, involving an integration of CMOS or other semiconductor-based technology and microfluidics. In one implementation, various components relating to the generation of electric and/or magnetic fields are implemented on an IC chip that is fabricated using standard protocols. The generated electric and/or magnetic fields are used to manipulate and/or detect one or more dielectric and/or magnetic particles and distinguish different types of particles. A microfluidic system is fabricated either directly on top of the IC chip, or as a separate entity that is then appropriately bonded to the IC chip, to facilitate the introduction and removal of cells in a biocompatible environment, or other particles/objects of interest suspended in a fluid. The patterned electric and/or magnetic fields generated by the IC chip can trap and move biological cells or other objects inside the microfluidic system. Electric and/or magnetic field generating components also may be controlled using signals of various frequencies so as to detect one or more cells, particles or objects of interest, and even the type of particle or object of interest, by measuring resonance characteristics associated with interactions between samples and one or more of the field-generating devices. Such systems may be employed in a variety of biological and medical related applications, including cell sorting and tissue assembly.

Description

Manipulation and/or detection to biological specimen and other object

Technical field

Relate generally to of the present invention is used for the method and apparatus via electromagnetic field manipulation, detection, imaging and/or identification particle or object.In various examples, integrated micro-system method and apparatus is disclosed, comprise by (for example using traditional semiconductor technology, Si, SiGe, CMOS, GaAs, InP) make and be configured to via electric field and/or magnetic field interaction guide, sensing, imaging and/or discern the electric field and/or the magnetic field generating apparatus of interested particle or object.In some example, such field generating apparatus and micro-fluidic system combine, and further make motion, sensing, imaging and/or the identification of interested particle or object become easy.

Background technology

In biological and medical science, it usually is useful can handling (for example, moving or guiding) biological specimen (for example, one or more cells) along the path of regulation.Manipulation based on the biosystem in magnetic field is a kind of method of using traditionally, finishing this task.In involving a kind of traditional embodiment in magnetic field, the little magnetic bead (magneticbead) with chemistry correction surface can be coupled to the target organism system, such as specific cell or little tissue.According to the type of the coating of given magnetic bead and relative size and target cell or little tissue of magnetic bead, magnetic bead can be adhered to the surface (coupled outside) of cell or little tissue, or by cell or little tissue picked-up (inner couplings)." magnetic bead is bonding " sample like this can be suspended in the main liquid (host liquid) then, constitutes " microfluid ", and the sample of the suspension in microfluid can be handled by using the external magnetic field then.Device based on this principle usually is called as " magnetic tweezer ", and (for example, DNA) confession research is used being used for collecting the little particle that floats on a liquid traditionally.

Because magnetic field and magnetic bead itself is biological typically compatible, so these process right and wrong are invasive and do not damage sample usually.Yet traditional magnetic tweezer can't provide the control respectively of a plurality of magnetic beads, because these devices typically only produce transportable single peak value; Therefore in microfluid, only can control single magnetic bead traditionally, or together the one group of magnetic bead closely of control simultaneously.

With the motion of the biological specimen that floats on a liquid, particle or other object with handle another relevant field and relate to the phenomenon that is called as dielectrophoresis.When inducing dipole on the particle that inhomogeneous field is floating on a liquid, dielectrophoresis takes place.Power subsequently on dipole is moved particle at the minimum value or the maximal value place of electric field.Almost any particle, without any special preparation, when being exposed to this suitable earth electric field, can be by using dielectrophoresis to be collected or moving.This operation that is based on electric field surpasses the advantage of above-mentioned operation based on magnetic field, because the latter requires with marked by magnetic bead biological specimen or other interested object.Yet the potential shortcoming of dielectrophoresis is that stronger electric field may damage cell, particle or other interested object under some environment relatively.

Can in medical diagnosis and life science, realize various application, with the motion of biological specimen with handle another relevant field and be called as " miniflow ".Miniflow is learned by the environment that provides support and keep the homeostatic microscale bio-compatible of physiology of cell and tissue to come at the inhibition of little biological specimen and/or flow.Micro-fluidic system can be configured for better simply relatively chamber or the liquid storage pool (" bathtub ") that maintenance comprises the liquid of interested cell/biological specimen; Replacedly, such system can have much complicated device, that comprise a plurality of pipelines or passage, and cell, particle or other interested object can flow therein.By control fluid flowing in microscale channel, a spot of sample can be by the route guidance to want in micro-fluidic system.Such as the such various microfluidic devices of valve, filtrator, mixer and dispenser, combine with microchannel in more complicated micro-fluidic system, can make the bioanalysis of the exquisiteness on the microscale become easy.Owing to allow to carry out many soft lithographic technology of duplicating for making in batches, even cost economy is also thought in the manufacturing of traditional micro-fluidic system of some complexity usually.

Yet in case made, traditional micro-fluidic system (particularly more complicated system) does not provide tangible flexibility ratio, and does not have enough programmability and controllability especially.Especially, be used in fixing passage and the valve that is manufactured with specific number and setting such as traditional micro-fluidic system of the such analysis operation of cytological classification.The operation control cell of valve flow in the passage, thus with they classification.The function of system is based on the statistical method of distinguishing between the cell of relatively large number usually, rather than cell of a subseries.Because the setting of passage and valve determines that during micro-fluidic system is made each system is designed to specific operation, and the basic structure of typically not revising it then can't be used in different processes.

Integrated circuit (IC) technology was the most great one of the technology that enables in a last century.I C technology is based on uses various semiconductor materials (for example, silicon Si, SiGe SiGe, gallium arsenide GaAs, indium phosphide InP or the like) to implement various electronic units and circuit.Perhaps one of most popular example of IC technology is CMOS (complementary metal oxide semiconductor (CMOS)) technology that is used for making silicon integrated circuit.

CMOS technology make now become the conventional part of daily life, calculating and communications applications such as the advanced person of personal computer, cell phone and wireless network or the like for some examples become possibility.The growth of computing machine and communication industry relies on electronics and continuous progress in technology relevant aspect the speed of the size that reduces of silicon integrated circuit and raising widely, and its trend is quantized by the Moore law usually.Current, silicon CMOS chip can surpass 100,000,000 transistors and with the speed operation of several GHz in little comprising in the structure of 90 nanometers.The CMOS micro-fabrication technology is ripe widely in nearly decades, makes silicon integrated circuit very cheap.Yet although a plurality of advantages are arranged, the IC technology of CMOS or any other based semiconductor all is not widely used in the structure that (that is transnormal data processing function) implements to be used for handling and characterizing such as sample such biologic applications.

Summary of the invention

The applicant recognize and the technology of seeing the integrated circuit based semiconductor (for example, Si, SiGe, GaAs, InP or the like), CMOS technology particularly, the feasible basis that is used to realize to handle with the system and method for characterising biological material and interested other object is provided.And the applicant recognizes and sees, by combined the technology of CMOS or other based semiconductor and miniflow, can realize various useful and strong, relevant with other material with biomaterial method and apparatus.

In view of foregoing, the various embodiment of present disclosure comprise the technology and combining that miniflow is learned of CMOS or other based semiconductor, relate to or multinomial method and apparatus of the manipulation, detection, imaging, sign, classification and the assembling that are used for biomaterial or other material on microscale.

For example, embodiment is at the combined IC/ miniflow commingled system of the bio-compatibility of the ability of integrated circuit (IC) chip and micro-fluidic system.Aspect of present embodiment, relate to the electric field of such commingled system and/or the various parts of magnetic field generation and use standard agreement (for example, to be implemented on the IC chip of CMOS) making in chip manufacturing factory.On the other hand, a generation parts itself can be formed by using the standard CMOS agreement, therefore without any need for micromachining technology (for example such in microelectron mechanical structure or MEMS embodiment).The electric field and/or the magnetic field that generate from such IC chip can be used for handling and/or detecting one or more insulating particles and/or magnetic particle, and distinguish dissimilar particles.

Especially, in one embodiment, the array of little electromagnet or " little coil " is implemented on the IC chip, and is configured to produce controllable, as to have the pattern of space and/or time magnetic field.On the one hand, the IC chip also can comprise programmable digital switching network and one or more current source, and it is configured to the electric current in each little coil of array of controls independently, so that create the magnetic field of the pattern that has space and/or time.On the other hand, the IC chip also can comprise humidity control system, to realize the bio-compatibility of commingled system.

In another embodiment, the array of microelectrode or " microtrabeculae " is implemented on the IC chip, and is configured to produce controllable, as to have the pattern of space and/or time electric field, so that handle interested particle according to the dielectrophoresis principle.On the one hand, the IC chip also can comprise programmable digital switching network and one or more voltage source, and it is configured to be controlled at independently the voltage on each microtrabeculae of array, so that create the electric field of the pattern that has space and/or time.As the embodiment of front, on the other hand, the IC chip also can comprise humidity control system, to realize the bio-compatibility of commingled system.

In another embodiment, the array of little coil of implementing on the IC chip can be configured to produce the electric field and/or the magnetic field of controllable, as to have space and/or time pattern.On the one hand, the IC chip also can comprise programmable digital switching network, together with one or more current sources and one or more voltage sources, be configured to be controlled at independently electric current and voltage in each little coil of array, so that create the magnetic field and the electric field of the pattern that has space and/or time.At present embodiment on the other hand, when being coupled with voltage on little coil, little coil plays the effect of microtrabeculae effectively, is used for thus handling interested particle according to the dielectrophoresis principle as former embodiment.Again, also can comprise humidity control system, to realize the bio-compatibility of commingled system according to the IC chip of present embodiment.

Combine with any above-mentioned embodiment that relates to electric field and/or magnetic field generation, another embodiment according to present disclosure, micro-fluidic system can directly be produced on the top of IC chip, or as the entity that separates, promptly suitably be combined on the IC chip, so that under the environment of bio-compatible, introduce and remove cell, or be suspended in interested other the particle/object in the fluid.Like this, can collect and move biological cell or other object in micro-fluidic system by IC chip electric field that generate, patterning and/or magnetic field.

Other embodiment of present disclosure is at the sensing/formation method and the equipment of one of other setting that utilizes above magnetic field and/or the electric field of introducing based on IC to generate array or magnetic field and/or electric field generating apparatus.For example, various aspects at these sensings embodiment, micro-coil array, other setting of a micro-pillar array or a generating apparatus (for example, consult on November 5th, 2002 and submit to, exercise question is " System and Method for Capturing and PositioningParticles; " PCT patented claim No.PCT/US02/36280, the various structures of describing among International Publication No. WO 03/039753 A1) can be by using the signal Be Controlled of various frequencies, so that can detect one or more cells by measuring the resonance characteristics related with the interaction between sample and one or more generating apparatus, particle or interested object, and even the type of particle or interested object.

In certain embodiments, radio frequency (RF) signal is utilized to be convenient to detection, imaging and/or identification.As one of the principle on the basis of these RF embodiment be the RF field energy enough almost with any (biological or other) but that under the RF signal frequency, conduct electricity or the electric or magnetic polarization particle interaction.Therefore, in these RF sensings embodiment, the interaction between near the object RF field and the RF field can be utilized to determine the position of one or more interested objects, so that realize the imaging of object.Like this, can be configured to detect and imaging biological cell, particle and interested object via electricity/magnetic device as disclosed herein, based semiconductor/miniflow commingled system and method, and need not depend on chemicals or optical technology by use RF signal is pure.According to such RF imaging technique, can comprise feedback control mechanism according to the various embodiments of the commingled system of present disclosure, interested thus sample can be handled according to the sample image that obtains.

In some aspects, RF technology disclosed herein singly can not be used in and detect and particles for imaging, and can also be used to discerning dissimilar particles/interested object.Such identification for example can be by measure R F field spectral response/on frequency range widely particle interaction and these responses and the various materials known characteristic that depends on frequency in electromagnetic field compared finishes.On the other hand, RF technology disclosed herein also can be used to carry out the magnetic resonance local measurement of (comprising ferromagnetic resonance) under the uniform magnetic field that is applied on sample or the interested object, thus (for example according to spin, electron spin resonance or " ESR ") or the material of the characteristic oscillation frequency recognition sample of magnetic domain (for example, nuclear magnetic resonance or " NMR ").Therefore, the method and apparatus according to the various embodiment of present disclosure can be utilized to implement effectively magnetic resonance imaging (MRI) system on chip.

In view of manipulation, detection, imaging and recognition technology discussed above and that discuss in more detail below, the applicant recognizes and sees that based semiconductor disclosed herein/miniflow commingled system and method make the various novel biological medicine and research and other application of systems biology be more prone to.

For example, another embodiment of present disclosure is at by utilizing IC/ miniflow mixed method and equipment and as the RF sensing/formation method of above introduction and the cytological classification method and apparatus of equipment.On the one hand, the cytological classification method and apparatus according to present embodiment makes accurate identification in molecule degree ground and cytological classification quick, pinpoint accuracy become easy.Especially, biological cell can with the accuracy of extra-high-speed and with the identification of the accuracy of molecule degree by each ground classification.The separation of specific (for example " rareness ") cell type or pathogen (stem cell that for example is used for the marrow-reconstitution operation of cancer patient) has been convenient in the classification of accuracy like this, is used for clinical practice.The classification of accuracy also has been convenient to analyze the statistics of tissue and estimate every kind of cell type discretely like this, rather than collects the gene expression data of related organization from the set of different cell types.

Another embodiment of the present invention is at the method and apparatus that is used to assemble microscale engineering tissue (engineered tissues).Aspect of present embodiment, be configured to and carry out microscale tissue assembling (for example, the artificial tissue that distributes from the abnormal shape of cell can once assemble a cell) with accurate control cell colony and space distribution based on the two-dimentional cell harvesting array of IC/ miniflow commingled system.Such technology representative according to an embodiment of present disclosure is used for developing the novel new method in external chemical examination, be used to study the communication network between different cell types, efficacy of drugs and be used for basic physiology research in standardized repeatably mode.

IC/ miniflow commingled system according to the based semiconductor of the various embodiment of present disclosure has several important techniques advantages.At first, the commingled system of based semiconductor/miniflow can be manufactured with high yield in the mode of suitable cost economy by using mature C MOS technology and being used to form micro-fluidic system cheap lithography technique partly.The system that such CMOS implements can be made on the size quite little and suitably be packaged into and can resist all-environment harm.Advanced low-power integrated-circuit technology also has been convenient to make battery powered device.In view of above-mentioned content, such system can be made into coarse disposable device, and can be used in various application, be included in to avoid when using traditional method and apparatus, harmful and/or urgent situation potentially.For example, can be easily and be used in the emergency medical situation effectively according to the commingled systems various embodiment, little, cheap, battery powered, coarse of present disclosure, by using saliva, breathing, sweat or blood sample quick check individual's health.Such system also can be used in material harmful on detection of biological under the given environment.

In addition, handle with traditional magnetic of using simple magnetic tweezer or external magnets, or compare with traditional dielectrophoresis technology, according to the commingled system of the based semiconductor/miniflow of present disclosure and method can be in large quantities and be easy to, mode accurate and quick control handle single or multiple biological cells, particle or other interested objects.And, provide significant flexibility according to the IC/ miniflow commingled system and the method for the based semiconductor of the various embodiment of present disclosure compared with traditional micro-fluidic system.Especially, slightly more complicated traditional micro-fluidic system is controlled biological specimen in the fixing channel network by using predetermined valve; Therefore, the different different specific micro-fluidic system of action need.On the contrary, can carry out various and exquisite cell/particle manipulation operations according to the commingled system and the method for based semiconductor/miniflow of the various embodiment of present disclosure, and not necessarily need complicated micro-fluidic system structure.

For example, in one embodiment, can implement by using according to the commingled system able to programme of present disclosure with relative better simply micro-fluidic system system's merging of the based semiconductor of the electromagnetic field that provides programmable and can independently control, that only have a single chamber (" bathtub ").In the present embodiment, cell almost can move through chamber at the computer control lower edge of electromagnetic field any paths.Like this, the topology that is used for " the virtual microscale pipe system " of interested sample can be changed at various operations neatly based on the programmability that is provided by computer control.This is for handle the extremely strong instrument that provides for accurate cell/object in better simply and more exquisite relatively operation.

In a word,, comprising at an equipment according to an embodiment of present disclosure: the field that a plurality of CMOS make generates parts; Near the micro-fluidic system field that a plurality of CMOS make generates parts, that be configured to comprise fluid; And at least one controller, be configured to control the field generation parts that a plurality of CMOS make, have enough intensity and the interactional electric field of at least one sample or the magnetic field that are suspended in the fluid to generate at least one.

, comprise from the field that a plurality of CMOS make generating the behavior that parts generate at least one electric field or magnetic field at a kind of method according to another embodiment of present disclosure; Described at least one electric field or magnetic field have enough intensity and interact with at least one sample that is suspended in the fluid, and this fluid is comprised in field that a plurality of CMOS make and generates near the parts the micro-fluidic system.

Should be appreciated that in more detail below all combinations of the above-mentioned notion discussed and additional concepts are intended as the part of theme of the present invention disclosed herein.Especially, all combinations of the claimed subject content that occurs at the end of present disclosure are intended as the part of subject content of the present invention disclosed herein.

Description of drawings

Fig. 1 is the block diagram that illustrates according to total looks of the various parts of the commingled system of based semiconductor/miniflow of an embodiment of present disclosure;

Fig. 2 illustrates the physics setting according to the example of the parts of a commingled system embodiment, shown in Figure 1 of present disclosure;

Fig. 3 (a)-(d) illustrates according to a little electromagnet line matrix embodiment of present disclosure, that an example of the magnetic field generation parts that can be included in the commingled system shown in Fig. 1 and 2 is provided;

Fig. 4 be according to an embodiment of present disclosure, also can be used as the synoptic diagram that magnetic field in the commingled system shown in Fig. 1 and 2 generates " annular collector " of parts;

Fig. 5 (a) and (b) be according to an embodiment of present disclosure, provide the electric field that can be included in the commingled system shown in Fig. 1 and 2 to generate the micro-pillar array of an example of parts;

Fig. 6 (a) is according to a conceptual perspective view embodiment of present disclosure, that can generate the micro-coil array of parts as the field in the commingled system shown in Fig. 1 and 2;

Fig. 6 (b) is the conceptual view according to (top) top view of the part of an array embodiment of present disclosure, shown in Fig. 6 (a), the part of the microchannel by comprising liquid is observed array downwards, and wherein suspending in liquid comprises the exemplary sample that is attached to the magnetic bead on the cell;

Fig. 7 (a) and 7 (b) illustrate skeleton view and the exploded view according to the little coil of a multilayer embodiment of present disclosure, that can be utilized respectively in the array of Fig. 6 (a) and 6 (b);

Fig. 8 conceptually illustrates the vertical layer structure according to the part of the CMOS IC chip of an embodiment of present disclosure, and it illustrates the Fig. 7 (a) relevant with layer with other characteristic of chip and the little loop construction of multilayer of 7 (b);

Fig. 9 illustrates according to an exemplary magnetic field pattern embodiment of present disclosure, when electric current flows through little coil on be similar to the little coil of those multilayers shown in Fig. 7 and 8;

Figure 10 conceptually illustrate according to an embodiment of present disclosure, Fig. 6 (a) and (b) shown in two adjacent little coils of array, wherein the electric current that equates basically flows through little coil, to generate two magnetic field peak values that equate basically;

Figure 11 (a)-(e) illustrate according to an embodiment of present disclosure, at five kinds of example scenario of adjacent little coil of Figure 10, in each coil, have the size of current of variation and the magnetic field that finally obtains of direction and generation;

Figure 12 is the every kind of situation that illustrates for five kinds of example scenario shown in Figure 11 (a)-(e), the size of current in each coil and the figure of direction;

Figure 13 illustrate according to an embodiment of present disclosure, be similar to micro-coil array shown in Fig. 6 (a) and related with this array various control assemblies;

Figure 14 illustrate according to an embodiment of present disclosure, parts is various interconnected in the first quartile of the array of Figure 13;

Figure 15 illustrates the content according to the little coil crosspoint in a little coil unit embodiment of present disclosure, that be included in first quartile shown in Figure 14;

Figure 16 illustrates according to a details embodiment of present disclosure, that provide current to the current source of first quartile shown in Figure 14;

Figure 17 illustrates according to a setting embodiment of present disclosure, that be used to be convenient to RF/ detection part pattern detection, formation " frequency-locked loop ";

Figure 18 illustrates the further details according to a phase detector embodiment of present disclosure, in frequency-locked loop shown in Figure 17;

Figure 19 illustrate according to an embodiment of present disclosure, in the further details of the phase comparator of phase detector shown in Figure 180;

Figure 20 illustrates the setting according to the replacement of RF/ detection part another embodiment of present disclosure, that be used to be convenient to pattern detection;

Figure 21 illustrates a setting embodiment, temperature controlling unit according to present disclosure:

Figure 22-26 illustrates according to a various treatment steps embodiment of present disclosure, that comprise when making based on the micro-fluidic system of polyimide as the part of commingled system;

Figure 27-32 illustrates according to a various treatment steps embodiment of present disclosure, that involve when making micro-fluidic system based on the patterning of ultraviolet solidifiable epoxy resin;

Figure 33-38 illustrates according to a various treatment steps embodiment of present disclosure, that comprise when making micro-fluidic system based on the soft lithographic technology;

Figure 39 (a)-(d) illustrate according to an embodiment of present disclosure, via as the exemplary of the cell detection of the above RF detection technology of discussing in conjunction with Figure 17-20;

Figure 40 illustrate according to an embodiment of present disclosure, based on the cytological classification equipment of the commingled system of Fig. 1 and 2; And

Figure 41-43 illustrates the assembly method of organizing according to a commingled system embodiment of present disclosure, that use Fig. 1 and 2.

Embodiment

Be below be used for handling, detection, imaging, sign, classification and assembling is biological or one of other material or multinomial, according to the more detailed description of the embodiment of the method and apparatus of present disclosure and each conception of species relevant with it.Should be appreciated that above introduction and in more detail below any way of the various aspects of main topic of discussion content in can be in many ways be implemented because subject content is not limited to any concrete mode of embodiment.Specific embodiment and examples of applications mainly are provided for illustrative purposes.

I. The total looks of system

An embodiment of present disclosure is at the bio-compatibility of the ability of microelectronic device and micro-fluidic system commingled system combined, based semiconductor/miniflow.In some example below, for the purpose of illustrating, the microelectronic device of commingled system part is implemented in CMOS technology.Yet, should be appreciated that present disclosure and be not intended to limit this respect, because the technology of other based semiconductor can be utilized to implement the microelectronic device various aspects partly of system discussed here.

Fig. 1 is the block diagram of total looks of various parts that the commingled system 100 of based semiconductor/miniflow is shown, and Fig. 2 illustrates the exemplary physics setting according to a parts embodiment, that be used for such system of present disclosure.As illustrated in fig. 1 and 2, commingled system 100 comprises micro-fluidic system 300, is used for keeping comprising the interested object liquid of (after this being called " sample ").Commingled system also comprises a plurality of other parts, comprises that electric field and/or magnetic field generate parts 200, a control assembly 400 and temperature controlling unit 500.Usually, these other parts can be utilized to be convenient to via electric field and/or magnetic field manipulation (for example, collect and/or move), detection, imaging and/or the identification of sample, regulate the biological specimen of environmental baseline (for example temperature) comprising needs.

Aspect of present embodiment, as shown in Figure 2, some of commingled system 100 or these all other parts can be implemented as one or more integrated circuit (IC) chip 102 by using various semiconductor fabrications.For example, Fig. 2 illustrates various generation parts 200, a control assembly 400 and temperature components 500 and can be fabricated on according to any technology in the various semiconductor fabrications on the semiconductor chip 104, to form IC chip 102.As describing in the above and discussing in more detail below, an exemplary of such IC chip can be made by using the standard CMOS agreement.IC chip 102 further can also be installed on the enclosed chip 110, and closing line 106 and contact site (for example, pin) 108 can be utilized to be convenient to and being electrically connected of IC chip 102.Among the embodiment who further discusses, a control assembly 440 also can comprise various parts, is used for being convenient to IC chip 102 with from the data of IC chip 102 and the radio communication of control signal below.

Fig. 1 and 2 also illustrates one or more processors 600, is configured to the various parts of control commingled system 100, so that for the manipulation that is comprised in the sample in (or flowing through) micro-fluidic system 300.One or more processors 600 also can be configured to carries out various signal processing functions, so that one or multinomial detection, imaging and identification of sample.Be to be understood that, in various configurations, described one or more processor 600 can be implemented as and commingled system 100 separated components, and randomly be placed with away from commingled system, (for example, various traditional computing equipments are coupled to commingled system via one or more contact sites 108 or via radio communication) as shown in Figure 2.Alternatively, some or all functional processor can be implemented by the element that combines with other parts in one or more IC chips 102 of a part that forms commingled system 100.

In the commingled system 100 according to an embodiment, micro-fluidic system 300 can be configured to be used to keep comprise the better simply relatively chamber or the liquid storage pool of the liquid of interested sample.For example, as usually illustrating on Fig. 1 and 2, the miniflow liquid storage pool with substantial rectangular volume can comprise

access pipeline

302 and 304, so that fluid flows into and the outflow liquid storage pool.Alternatively, micro-fluidic system can have more complicated setting, comprises a plurality of pipelines or passage that the liquid flow that comprises sample is wherein arranged, and is used to guide mobile various parts (for example valve, mixer).In various embodiments,, just micro-fluidic system 300 can be produced on the top of the IC chip 102 that comprises other system unit, to form commingled system 100 in case finish semiconductor fabrication; Alternatively, micro-fluidic system 300 can be made (for example using the soft lithography technology) separatedly, and is attached to subsequently on the one or more IC chips that comprise other system unit, to form commingled system 100.Discuss in the further details of the relevant micro-fluidic system 300 V joint below.

Others at embodiment shown in Figure 1, the electric field of commingled system 100 and/or magnetic field generate parts 200 and can be placed with various settings with respect to micro-fluidic system 300, so that in the field that generates and be comprised in interaction between the sample in (or flowing through) micro-fluidic system.In various embodiments, a generation parts 200 can be set near the micro-fluidic system along one or more physical boundaries of micro-fluidic system, and is configured to allow the edge to interact about the field-sample of one or more Spatial Dimensions of micro-fluidic system.

For example, in one embodiment, as shown in Figure 2, micro-fluidic system 300 can be configured to the liquid storage pool of the substantial rectangular shape on the IC chip 102 of the two-dimensional array that comprises field generation parts 200, and its midfield generating unit part 200 is set near the liquid storage pool and is arranged essentially parallel in the plane of liquid storage pool bottom surface.The operation of common sample along two dimensions (the x-y axle by Fig. 2 is represented) that limit the plane be parallel to the liquid storage pool bottom surface has been convenient in such setting.In another embodiment, generating parts can be alternatively or be set up along one or more sides of this liquid storage pool in addition, so that along the operation of the sample of the third dimension (the z axle by Fig. 2 is represented) that is transverse to (for example, perpendicular to) liquid storage pool bottom surface.In a further embodiment, liquid storage pool can by " sandwich " be comprised in respectively be set at above the liquid storage pool and below the IC chip in the field generate between two arrays of parts.In such setting, a plurality of arrays that generate parts can be controlled such that the Three dimensional steerable that can finish sample.In addition, a generation parts can be so that the rotation of sample with respect to the various settings of micro-fluidic system.

Be to be understood that, above-mentioned exemplary the setting mainly is provided to be used for explanation, and the various settings of a micro-fluidic system and a generation parts (comprise the straight line or the two-dimensional array that generate parts, or discrete field generates other setting of parts) according to other embodiment, expection provides the multidimensional of sample to handle.In a word, according to each conception of species discussed here, generating by the field under the electric field and/or the control of CALCULATION OF MAGNETIC FIELD INDUCED AROUND machine that parts 200 generate, interested sample can move through micro-fluidic system along any paths almost, be collected or be maintained at certain location, and be rotated in some cases.Like this, being used for topology interested sample, " virtual microscale pipe system " can for example be changed according to available programmability and computer control by processor 600 neatly, is used for various operations.This handles the extremely strong instrument that provides for accurate cell/object in better simply and more exquisite relatively operation.

In the various embodiment of the commingled system shown in Fig. 1 and 2 100, generate parts 200 and can be configured to generate electric field, magnetic field or the two.For example, in one embodiment, generate parts to be configured and to be operated, with generation extend in the micro-fluidic system, controllable, in the space and/or the magnetic field that changes on the time.The magnetic field of Sheng Chenging interacts with the magnetic sample that is suspended in the micro-fluidic system like this, and sample wherein includes but not limited to, is attached to the biological cell (" in conjunction with the cell (bead-bound cells) of magnetic bead ") on the magnetic bead.With respect to biological specimen, it should be noted that magnetic field do not damage cell; But, as discussed above, be normally used technology via the cell operation and the identification in magnetic field, it applies the magnetic bead of (ligand-coated) and molecule degree ground identification biological cell by specific, aglucon.As following further discussion, the interaction between space and/or magnetic field that changes on the time and the cell that combines magnetic bead or other magnetic sample enables single or multiple magnetic sample collections, conveying, detection and imaging.

Example that magnetic field in the commingled system 100 shown in Fig. 1 and 2 generates parts 200 be can be included in and two-dimentional little electromagnet line matrix shown in Fig. 3 (a)-(d) and one or more " annular collector " as shown in Figure 4 included but not limited to.These example components were submitted on November 5th, 2002, exercise question is the PCT patented claim No.PCT/US 02/36280 of " System and Method for Capturing and PositioningParticles ", is gone through among International Publication No. WO 03/039753 A1.

Fig. 3 (a) is the synoptic diagram of little electromagnet line matrix 200A.According to an embodiment, matrix (for example comprises straight basically conductor, the gold or other metal wire or rail) top layer 202 and bottom 204, wherein every layer by insulation course 206 (for example, polyimide) covers, the conductor of equivalent layer is set up (for example, the conductor of top layer is perpendicular to the conductor of bottom) in the crosscut mode.In different embodiments, this structure can be fabricated on the various substrates, and an example of this substrate comprises sapphire substrate.Fig. 3 (b) illustrates the micrograph of the line matrix of the such making that comprises electric connecting wire, wherein represents in the legend in the lower right corner of figure at the exemplary dimensions of the device of the making that illustrates.Fig. 3 (c) illustrates the part of the amplification of the device shown in Fig. 3 (b), and it is basically corresponding to the explanation of the notion of Fig. 3 (a).At last, Fig. 3 (d) is the micrograph of the sectional view of device, shows two layers of vertical conductor/insulation body structure.

In a embodiment based on the line matrix shown in Fig. 3 (a)-(d), each conductor in the line matrix (or alternatively, predetermined conductor group) can be connected to controllable current source (following further discussion), so that all conductors (or conductor group) can have independently electric current.By modulating the amplitude of electric current in the conductor independently, (for example) can produce various dynamic magnetic field patterns near the line matrix.For example, electric current can be controlled such that the line matrix can create continuously the single magnetic field peak value that moves, a plurality of peak values that each peak value is independently controlled, or the variation magnetic field, so that rotation or reverse target sample.

Fig. 4 is the synoptic diagram that also can be used as " annular collector " 208 of the magnetic field generation parts in the commingled system shown in Fig. 1 and 2.Annular collector is the conductor (for example, being set at sapphire or other on-chip gold thread or rail) that is set at on-chip single circular carries current basically, has insulation course at the top.Because electric current flows through round conductor, so generate magnetic field from annular collector; In an example, in the circular rings of diameter with about 5 microns (μ m), flow through 30 milliamperes of magnetic fields that (mA) electric current can produce about 10 Gausses of conductor, corresponding to the magnetic force (for example, it surpasses the power that is enough to attract and collect the bacterium that adheres to magnetic bead) of about 10 skin oxen (pN).Such annular collector can be provided with various configurations with respect to micro-fluidic system, comprises the one dimension or the two-dimensional array of annular collector.

Another example that can be used as the device of the magnetic field generation parts in the commingled system shown in Fig. 1 and 2 comprises magnet microscale, that be configured to coil or " little coil ".Comprise ferromagnetic core and some examples by using little coil that micromachining technology makes in U.S. Patent No. 6,355,491 and 6,716,642 and international application published No.WO 00/54882 in provide.Another example that generates parts according to the magnetic field of one embodiment of the present of invention comprises CMOS micro-coil array and relevant control circuit.The further details of such CMOS micro-coil array II joint is below discussed.

Be to be understood that, for according to present disclosure, go up almost any commingled system 100 of the microelectronic device part of controllable variable magnetic field based on being configured to be created on space and/or time, parallel embodiment can by use be used to generate controllable, in the space and/or the configuration of the combination of temporal variable electric field or variable magnetic field and variable electric field be implemented.

For example, in one embodiment, the field of the commingled system shown in Fig. 1 and 2 generates the array that parts 200 can comprise microelectrode or " microtrabeculae ", and it is configured to generate controllable electric field, is used for handling interested object according to the principle of dielectrophoresis.Fig. 5 (a) and the example of such micro-pillar array 210 (b) is shown; Fig. 5 (a) illustrates the micrograph of the top view of the micro-pillar array that comprises making electric connecting wire, such, wherein 15 microns (μ m), represent in the legend on the left side of figure at the exemplary dimensions of the device of the making of being drawn, and Fig. 5 (b) illustrates the skeleton view of amplification of the exemplary array of Fig. 5 (a), shows the two-dimensional arrangements of 5 row and 5 capable microtrabeculaes.

As discussed above, dielectrophoresis takes place when the dipole on the particle of inhomogeneous field induction suspending in liquid.Be added to minimum value or maximal value place that the power of dipole is moved particle in electric field subsequently.Need not any special preparation, almost any particle when it is exposed in the suitable internal field, just can be collected or be moved by using dielectrophoresis.Like this,, be applicable to the electric field of this task with generation, can handle the one or more interested sample that in micro-fluidic system 300, floats on a liquid by operation micro-pillar array 210 according to an embodiment.

More particularly, based on Fig. 5 (a) with among the embodiment of the micro-pillar array 210 (b), each microtrabeculae in array (or alternatively, predetermined microtrabeculae group) can be connected to controllable voltage source (following further discussion), like this, all microtrabeculaes (or microtrabeculae group) can have independently voltage potential thereon.By being modulated at the amplitude of the voltage on the corresponding microtrabeculae independently, can near micro-pillar array 210, (for example, produce various electric field patterns in the above), so that handle the one or more interested sample that is comprised in the micro-fluidic system.In order to be provided for the ground of each microtrabeculae current potential, an exemplary geometric structure is included in micro-pillar array next door and top (for example, on the lower surface of miniflow chamber) making ground plane, and like this, the line of electric force of nearly all generation points to same direction.Alternatively, can generate electric field maximum, avoid ground plane thus by different voltage potential (for example, positive with negative is connected) is applied to (for example, adjacent) microtrabeculaes different in array.

In another embodiment, that micro-coil array can be configured to produce is controllable, have the electric field and/or the magnetic field of pattern in the space and/or on the time.More particularly, in the embodiment of further discussing in the II joint below, each voltage that can control independently can be applied on little coil of micro-coil array, like this, each little loop construction performance basically image pattern 5 (a) and (b) shown in the microtrabeculae of micro-pillar array 210, that is, by generate can with the interactional electric field of sample that is comprised in the micro-fluidic system.According to an aspect of present embodiment, corresponding controllable independently electric current also can be applied on little coil of micro-coil array, so as additionally to generate can with the interactional magnetic field of magnetic sample that is comprised in the micro-fluidic system.These and other type, can be used in based on electric field or based on the embodiment of electric field/magnetic field and to relate to the various application that merged manipulation, sensing and imaging system that microelectronics and miniflow learn.

With shown in Figure 1, according to some embodiment of the present invention, the field control assembly 400 of commingled system 100 can comprise one or more current sources 420, generates magnetic field so that generate parts by magnetic field as mentioned above.Similarly, according to other embodiment of the present invention, a control assembly is all right-or alternatively-comprise generating electric field so that generate parts by one or more voltage sources 440 by electric field.

Usually, no matter a control assembly 400 comprises one or more current sources 420, one or more voltage source 440, or the two, according to an embodiment, the field control assembly also comprises various exchanges or multiple connection parts 460, so that electric current and/or voltage suitably are applied on each generation parts or the generation parts group.In the various embodiments that discuss in more detail below, exchange or multiple connection parts 460 (for example can be configured to the programmable digital switching network, under the control of one or more processors 600), like this, the output of one or more electric currents and/or voltage source is applied on the generating unit part of showing up with the independent controllable mode of regulation, is convenient to electric field and/or magnetic field that sample is handled, have pattern in the space and/or on the time so that set up.

Also as shown in Figure 1, various embodiment according to present disclosure, control assembly 400 also can comprise radio frequency (RF) parts and other detection part 480 that is coupled between generating unit part 200 on the scene and the one or more processor 600, is used for being convenient to the sample that or multinomial detection, imaging and sign are comprised in micro-fluidic system 300.Aspect different, the example of such RF/ detection part 480 can include but not limited to oscillator, frequency mixer and/or wave filter, they (for example, via exchange or multiple connection parts 460, under the control of one or more processors 600) moved, generate RF field and meter and be shown in the RF field that generated and interactional certain type signal between one or more interested sample to generate parts from the field.The concrete details III joint below that is used for the exemplary circuit embodiment of RF/ detection part 480 is further discussed.

In all fields, RF/ detection part 480 all is provided for purely pattern detection, imaging and the characterization technique based on electromagnetic field, and do not need to be harmful in the chemical element of interested sample, or heavy optical microscope.In any case, should be appreciated that according to comprising some technology of each conception of species disclosed herein pattern detection and imaging can be assisted with the sample of processing/aligning particular type chemically.

Usually, as known according to Maxwell equation, but the RF field energy enough with almost any (biological or other) that under the RF signal frequency, conduct electricity or the electric or magnetic polarization particle interaction.Therefore, in the various embodiment of present disclosure, the interaction between RF electric field and/or magnetic field and interested sample singly can not be utilized to mobile sample, can also determine the position (for example, being used to be convenient to imaging) of sample.And, from the RF field/the interact spectral response cause of sample can be used for discerning or characterizing the sample of different type or classification in some cases.

For example, the conduction sample has the ring current by the induction of RF field, and these electric currents produce their magnetic field again, and interacts consumingly with the field that is applied in.This is the basis (" squirrel-cage " rotor that for example, does not have electrical contacts) of the operation of traditional motor.This interaction can be used for mobile sample, and detects their existence.In the mechanism that discusses in more detail below, the magnetic polarization of sample changes near the inductance of the coil (for example, little coil of array) sample; Therefore, the decay of the vibration of magnetic polarization causes detectable loss in comprising the circuit of little coil.In another example, the electric polarization of sample causes the power that causes dielectrophoresis (DEP).This polarization can be via the change of the electric capacity between the electrode (microtrabeculae or little coil with the voltage that applies) of sample and electric field generating apparatus, and it is not detected not have the ground of dissipation, or by because the change of the decay that the electric polarization of vibrating in the sample causes is detected.Thereby above-mentioned example provides and can be used for detecting the position of sample and the various mechanism of imaging.

Based on such RF imaging technique, can introduce feedback control mechanism according to the various embodiments of the commingled system of present disclosure, interested thus sample can be handled according to the image of the sample that obtains.For example, in one embodiment, commingled system can (for example via one or more processors 600) be configured at first obtain being comprised in the image of the distribution of the sample in the micro-fluidic system able to programmely.After this, according to the distribution of imaging, one or more specific sample can be handled based on the algorithm of regulation.

Relevant with the RF field, each conception of species disclosed herein similarly can be utilized to identification and characterize interested sample.For example, the change that depends on frequency in the electric polarization of sample or magnetic polarization can be used for the type of recognition sample, has wherein used the knowledge from the behavior of relevant various materials in electromagnetic field of traditional solid state physics.These changes can be characterized in the frequency range of broadness.Therefore, in one embodiment, be applied in the RF frequency (or using more exquisite signal processing technology) of the signal on the generating unit part of showing up by scanning, the frequency response of sample (for example, absorption spectra) can be measured at the certain location place, and, can discern or characterize sample according to measured response.

About the RF field under the control of RF/ detection part 480 apply and another embodiment of the interactional sensing of field/sample in, the RF field can be used for local measurement and be applied to magnetic resonance in the uniform magnetic field on the sample.Especially, the spin or the magnetic domain of given sample vibrate with characteristic frequency, and this can be used for discerning the type of spin type or sample itself.The magnetic resonance type comprises ferromagnetic resonance (FMR) (little YIG ball can be used as magnetic bead, and wherein the YIG ball has single magnetic domain, rotates freely, because magnetic bead is spherical) on the GHz frequency.In addition, electron spin resonance (ESR) technology can be used for discerning the g factor of the spin of the involved origin (being sample) that characterizes them, and nuclear magnetic resonance (NMR), is used for discerning the g factor of nuclear spin.Therefore, according to principle discussed here, magnetic resonance imaging (MRI) system can be implemented on chip.

Though obviously do not illustrate on Fig. 1 and 2, but according to each embodiment, control assembly 400 also can comprise one or more analog to digitals (A/D) and digital-to-analog (D/A) converter so that between other control assembly and arrive with from the various data of IC chip 102 and communicating by letter of signal.The field control assembly also can comprise digital signal processing parts and signal amplification component, so that Signal Processing and conveying.And control assembly also can comprise transceiver and antenna so that to and from the radio communication of IC chip 102.In an example wireless embodiment, the ISM radio frequency band is (free, the non-commercial radio frequency band allows to be used for industry, scientific research and medical application) can be used in the radio communication between IC chip 102 and remote subscriber or control interface (for example, one or more processors 600).That current wireless receiving and dispatching machine technology allows is microminiaturized, lower powered transceiver sends and receives with high data rate (for example, several kbit of per second or Mbit), and this is enough to transmission information reliably to IC chip 102 with from IC chip 102 transmission information.

At last, Fig. 1 and 2 also illustrates, and commingled system 100 can comprise temperature controlling unit 500, so that the bio-compatibility of commingled system.For example, according to an embodiment, the temperature of system can be adjusted to or near specific temperature, so that the bio-compatibility of system and the sample of being studied.In an exemplary, temperature controlling unit can comprise one or more " monolithic (on chip) ", and temperature sensor 500A (for example, near micro-fluidic system 300, as shown in Figure 2) and " sheet outer (off chip) " temperature controller 500B (for example, being attached to thermoelectric (al) cooler or " TE " refrigeratory of enclosed chip 110).On the one hand, near the temperature of the IC chip of these one or more monolithic temperature sensor 500A sensings micro-fluidic system 300, and described one or more processor 600 compares measured temperature and reference temperature (for example, 37 ℃).Described one or more processor sends to the outer temperature controller 500B of sheet to suitable feedback control signal again successively, and it correspondingly heats or cools off whole substrate.Temperature controlling unit 500 IV joint is below further discussed.

Provide according to present disclosure pass through use total looks of commingled system of electromagnetic field manipulation, detection, imaging and sign sample after, set forth the more detailed description of each conception of species of the different part that relates to commingled system and some exemplary application below.

II. Micro-coil array

Fig. 6 (a) is according to a conceptual perspective view embodiment of present disclosure, that can be used as the micro-coil array 200B of the field generation parts 200 in the commingled system shown in Fig. 1 and 2 100.In the example of Fig. 6 (a), array 200B comprises five row and the substantially the same little coils 212 of the five-element.Though Fig. 6 (a) illustrates 5 * 5 micro-coil arrays, should be appreciated that the micro-coil array according to various embodiment of the present invention is not limited thereto, but can have little coil and different geometry arrangement of different numbers.

As above little electromagnet line matrix 200A in conjunction with Fig. 3 (a)-(d) discussion, be similar to the such micro-coil array 200B shown in Fig. 6 (a) and can be configured and be controlled to handle the magnetic sample that is comprised in the micro-fluidic system 300, comprise the cell that is coupled on the magnetic bead.Fig. 6 (b) illustrates the conceptual illustration of top (top) view of the part of the array 200B shown in Fig. 6 (a), the part of the micro-fluidic system 300 by comprising liquid 306 (for example, passage) observes array downwards, the exemplary sample 116 (that is the cell that, adheres to magnetic bead) that wherein in liquid, is suspending and comprising the magnetic bead 112 that is attached on the cell 114.Liquid 306 also can comprise one or more cells 114 that are not attached with magnetic bead.In one embodiment, in order to handle the cell 116 (or magnetic sample of other type) that adheres to magnetic bead, the little coil 212 of each of array 200B can be connected to the source of (via replacement part and multiple connection parts, as what further discuss below in conjunction with Figure 13) controllable electric current independently.Therefore, by controlling the amplitude of the electric current that flows through each little coil independently, can generate each magnetic field pattern near micro-coil array 200B, these magnetic field patterns are utilized to collect or the operating magnetic sample.

Compare with little electromagnet line matrix 200A, owing to the exemplary reason below some at least, micro-coil array 200B is normally more effective.At first, compare with in little electromagnet line matrix those in the field that generates in micro-coil array, by highland localization more, provides relative higher spatial resolution for the sample of collecting with carrying thus.Secondly, micro-coil array has meticulousr magnetic field control degree compared with little electromagnet line matrix, therefore can handle the more sample of big figure simultaneously; Especially, N * N micro-coil array can provide N effectively ²The local magnetic field of individual independently while is (based on N ²And N * N line matrix can only provide 2N independently simultaneously field (based on the individual independently electric current of 2N) individual independently electric current).The 3rd, as what discuss in more detail below, little coil is owing to its clearly inductance of definition provides the better platform that is used for the RF detection.The 4th and since the stray magnetic fields that causes of electrical lead usually in micro-coil array compared with in little electromagnet line matrix, being littler.

Relate to according to a problem in the design of the two-dimentional micro-coil array 200B of an embodiment of present disclosure and to be right after above the array and be parallel to the magnetic force that can generate on the plane of array.This plane is usually represented by x axle and y axle on Fig. 2 and 6 (a).Especially, the x-y component of the magnetic force that is generated by the corresponding little coil of array must be enough greatly, so that in reasonable range (for example, distance between two adjacent little hub of a spools, or " spacing " of array, as going up at Fig. 6 (a) by label 216 expressions) and in rational time (for example, 1 second or littler) overcome skin-friction force and fluid viscosity moves the magnetic sample (for example being attached to the biological cell on the magnetic bead) that is suspended in the fluid.Another design problem relates to the magnetic potential energy; Suppress simultaneously because the heat that the heat energy of sample causes (is for example shaken for the enough strong collection that keeps the magnetic sample, Brownian movement) and the diffusion, the magnetic potential that generates by corresponding little coil can be fully greater than the heat energy of sample (promptly, 3/2kT, wherein k be the graceful constant of bohr thatch and T is a sample temperature).Another design problem relate to perpendicular on the direction on the plane of array, along the magnetic force (the z axle shown in Fig. 6 (a) is in skeleton view, is actually the outside direction in plane of sensing figure) of the z axle shown in Fig. 2 and 6 (a).According to the technology and the method that are used for making micro-coil array; above array, one or more material layers can be arranged (for example; material layer of insulation, protection and/or bio-compatible or the like); these layers extend considerable distance along the direction that is parallel to the z axle above array; surpass this distance, the magnetic field that is generated is decline fast.

Remember above-mentioned problem, an embodiment of present disclosure is at by using traditional CMOS treatment process to be fabricated on semiconductor (for example, Si) on-chip micro-coil array.Aspect of present embodiment, each control assembly that comprises the control electronic installation that is used for micro-coil array is incorporated in and is made as CMOS IC chip with micro-coil array, the space and/or the time that are used for the sample manipulation for use in generation are gone up variable magnetic field, and RF field, so that pattern detection, imaging and sign.Especially, in exemplary, little coil itself is formed by using the standard CMOS agreement, therefore need not any micromachining technology (for example, as in micro electro mechanical structure or the MEMS embodiment).

More particularly, in order to solve above-indicated design problem, according to an embodiment, available many metal levels are utilized to allow to generate enough magnetic field intensitys in little coil configuration in CMOS manufacturing process, are enough to collect effectively and carry sample.Fig. 7 (a) and 7 (b) illustrate skeleton view and the exploded view according to exemplary three layers of little coil 212 of present embodiment respectively, and Fig. 8 conceptually illustrates the vertical layer structure of the part of CMOS IC chip 102, shown in it with other characteristic and three layers of relevant little coil of layer of total chip structure.Corresponding to also expression on Fig. 7 and 8 of the such z axle shown in Fig. 2 and 6 (a).Should be appreciated that the exemplary three layers of little loop construction shown in Fig. 7 and 8 mainly are provided for the purposes of explanation, can comprise different number zone of interest (for example, two or more) and/or have different overall shapes or geometric relationship according to little coil of other embodiment.Usually, according to various embodiment, be similar to the part (for example layer) that the such little coil shown in Fig. 7 and 8 can comprise the axial concentric apart of at least two of conductor ring.

Shown in Fig. 7 and 8, exemplary little coil 212 comprises conductor part or layer, that is: upper part 212A, center section 212B and the lower part 212C of three coilings.For the ease of accurately controlling to the space each magnetic sample in the micro-fluidic system that is comprised in micro-coil array 212 tops, each little coil is designed to generate single magnetic field peak value above little coil, to interact with sample.For example, conceptually illustrate as Fig. 8, when suitable electric current flows through little coil, be suspended in the liquid that is comprised in micro-fluidic system 300 the magnetic sample (for example, the cell that adheres to magnetic bead is shown in Fig. 6 (b)) attracted to above little coil 212 the peak value place, magnetic field that generates.In Fig. 8, represent with reference number 120 with the bottom of micro-fluidic system 300 or the distance between the bottom surface at the upper part 212A of little coil (as being fabricated on the structure of layering generally of IC chip 102).

Usually discuss as above, the work principle that is used for the micro-coil array 200B that the magnetic sample handles is to create and move one or more magnetic fields peak value by the electric current of each the little coil 212 that is modulated at array.For example, at first consider a little coil 212 of " connection " (that is, electric current flows through) array only (that is, shown in Figure 8 little coil); As shown in Figure 8, magnetic sample 116 attracted to the peak value place, magnetic field that is generated by little coil 212, therefore is collected in the center of little coil of the surface of IC chip 102.Near the magnetic field peak value that is generated, " collection power " is provided by following formula:

F = Vχl μ_{o} &dtri; B^{2}, - - - (1)

Wherein V is the volume of magnetic bead 112, and χ is the effective susceptibility of magnetic bead, μ ₀Be space permeability, and B is the magnetic field size that is generated.If this little coil subsequently by " shutoff " and simultaneously adjacent little coil of array be switched on, then the magnetic field peak value moves on to the center of adjacent little coil, thus magnetic bead is transported to new peak.

Proportional for generating the specific needed magnetic field B of collection power F with the inductance value of the electric current that flows through little coil and little coil; The inductance value of little coil is proportional with the number of turns of little coil and the size of little coil (diameter) again.Therefore, provide little coil design of higher relatively inductance value normally desirable, collect sample with the magnetic field that sufficient intensity is provided.Simultaneously, for the meticulous spatial resolution between the little coil that keeps array and the sample be convenient between adjacent little coil are carried, what wish usually is, have interval or spacing 216 and relative little diameter 214 between the less relatively coil of upper part 212A of little coil, shown in Fig. 6 (a).

Therefore, various aspects at present embodiment, the diameter of total number of turns of little coil and each coiling part is suitably selected, and so that suitable array pitch and suitable little coil inductance value to be provided, is convenient to sample collection and the conveying between little coil to generate enough magnetic field.For this reason, the vertical space of the little loop construction use of the multilayer shown in Fig. 7 and 8 in the CMOS of layering chip design obtains the bigger number of turns of each little coil, so that higher inductance value to be provided.Simultaneously, the number of turns distributes in the different level of little coil with partly, permission is different diameters on the different level/part at little coil, and this has been convenient to interval or spacing between the little coil between the adjacent coil, and effective little coil inductance value is provided simultaneously.

More particularly, in the exemplary little coil shown in Fig. 7 and 8, thereby the upper part 212A of the sample in the surface of the most close IC chip and the most close miniflow chamber can be made as the individual pen metallic conductor with less relatively diameter 214, and its size can be determined by the average-size of the sample that will be collected.In an exemplary, the diameter 214 of upper part 212A can be to be about 10-11 μ m; Should be appreciated that this diameter is greater than about 5 μ m usually owing to the restriction of present CMOS manufacturing technology.The diameter 214 of upper part 212A also can be selected according to the size of wanting generally of micro-coil array 200B and the spacing of wanting 216 to small part.Usually, in order to ensure the appropriate resolution between magnetic field adjacent, the interval between the upper part of adjacent little coil should be not less than the approximately diameter 214 of each upper part; This causes spacing 216 to be about the twice (again, the resolution that should be appreciated that the raising of array mainly is subjected to the resolution limit of manufacture process) of diameter 214.According to this general relation, in various embodiments, diameter 214 and spacing 216 scopes can be from several microns to tens microns, depend on the type of the sample of being considered and the application that is involved.

Shown in Fig. 7 (a) and 7 (b), the center section 212B of little coil and lower part 212C can have bigger diameter compared with upper part.On the one hand, the bigger diameter of center section and lower part is possible, because adjacent center section and the interval between the lower part at adjacent little coil of array can be less than the intervals between adjacent upper part, and do not sacrifice the resolution (that is, the resolution in the magnetic field that is generated is determined by metal layer at top greatly) in the magnetic field that is generated.Therefore, center section and lower part can comprise the number of turns of bigger number and/or bigger diameter compared with upper part usually, thus, provide higher relatively little coil inductance value.In addition, shown in Fig. 7 (a) and 7 (b), lower part 212C can comprise pad (pad) 228, so that being connected of little coil 212 and electric current (or voltage) source, as following further discussion.In an exemplary, each center section and lower part can comprise three conductor rings, wherein the diameter 220 of center section 212B can be the magnitude that is about 20-25 μ m, and the diameter 218 of lower part 212C can be the magnitude (the less relatively diameter of lower part allows to comprise pad 228) that is about 15-20 μ m.In other embodiments, the different conductor number of turns and/or different yardsticks can be used in the respective coil part, they can experience ground or determine according to Digital Simulation at the magnetic field of wanting of different application.

Referring now to IC vertical layer structure as shown in Figure 8, IC chip 102 comprises semiconductor-based lamella 104, sequentially makes three layer/

part

212C, 212B and the 212A of little coil 212 above the substrate layer at this.Each layer/

part

212C, 212B and 212A can form by deposition and the patterning such as the conducting metal of copper, gold or aluminium.A plurality of metal levels are separated mutually by the insulating material 112 that for example comprises Si oxide (SiO2) or other suitable electric dielectric material and are separated with other layer of IC chip.Three layer/

part

212C, 212B are in the same place by electric coupling with 212A, so that create continuous multilayer conductive loop (through hole 114 also is indicated on the skeleton view of Fig. 7 (a)) by the through hole 114 (for example, being made by tungsten) that extends through insulating material 112.

In one embodiment, the CMOS treatment technology (for example, the CMOS 0.18 μ m technology of Taiwan semiconductor manufacturing company) that is utilized to make vertical layer structure shown in Figure 8 produces the thickness 222 of about 1-3 μ m of upper metallization layer/part 212A.Upper metallization layer also can be patterned so that the x-y face of metallic conductor (that is, perpendicular to the plane of Fig. 8) line width also is about 1-3 μ m, so that the metallic conductor cross section of upper part 212A is from about 1 * 1 μ m ²To about 3 * 3 μ m ²(should be appreciated that the 0.18 μ m Design Rule according to TSMC, the line width of upper metallization layer-metal 6-can be small enough to 0.44 μ m).

For middle and

lower part

212B and 212C, the CMOS treatment technology can produce the thickness 224 of about 0.5 to 1 μ m of bottom and middle layer/part.These layers also can be patterned, so that also be about 0.5 to 1 μ m at the line width of x-y face, produce about 0.5 * 0.5 μ m ²To about 1 * 1 μ m ²The bottom and the metallic conductor cross section of center section.Distance 226 between metal level can be the magnitude (should be appreciated that the 0.18 μ m Design Rule according to TSMC, the distance between metal level can be small enough to 0.46 μ m) that is about 1 μ m.

According to above-mentioned overall yardstick, can obtain about 1 nanohenry (1nH) magnitude or higher little coil inductance value.By generally reducing the various yardsticks relevant with metallic conductor, wire circle can increase, and causes the inductance value up to 60 to 100 nanohenrys (60-100nH).Yet, should be appreciated that width when metallic conductor becomes more hour, the dead resistance of coil increases usually, and the electric current that maximum allows to flow through coil reduces usually, the intensity in the magnetic field that this final restriction can generate; Therefore, between coil dimension and field intensity, have actual trading off.

More generally, should be appreciated that vertical layer structure shown in Figure 8 is not limited to above-mentioned yardstick, or three metal levels; According to present CMOS manufacture craft, also be possible up to about seven metal levels.Therefore, again, three layers of little loop construction provides as just an example according to multiple possible little coil configuration of present disclosure.

Vertical layer structure as Fig. 8 is shown, and deposit passivation layer 116 on insulating material 112, and it for example can comprise silicon nitride or polyimide.At last,, dimethyl silicone polymer (PDMS) layer 118 is deposited on the passivation layer 116, is used as interface with micro-fluidic system 300 in order to ensure bio-compatibility.In various embodiments, can be the magnitude that is about 3-4 μ m in the upper metallization layer/part 212A and the distance between the interface of PDMS layer 118 and micro-fluidic system 300 of little coil.

According to the as above general structure of the little coil of CMOS of general introduction, can generate significant local magnetic field above little coil in each of array 200B, so that the manipulation sample.For the illustrative numerical range of magnetic field intensity and sample collection power is provided, consider to have total diameter of about 20 μ m and little loop construction of two layers of every layer of 4 wire circle.Exemplary little coil comprises having 1 * 1 μ m ²The aluminium conductor in average conductor cross section, wherein live width is 1 μ m, the gap between the adjacent cross-sectional area of conductor of given layer is 1 μ m, and the distance between two-layer is 1 μ m.The maximum current density of aluminium conductor is about 200mA/ μ m ²Therefore, exemplary little coil of being considered can support to flow through it, the maximum current of about 200mA.Fig. 9 is illustrated near the Distribution of Magnetic Field bottom surface of the micro-fluidic system that wherein has sample, on so little coil in the x-y plane at about 1 μ m place.As what on Fig. 9, see,, generate the significant magnetic field peak value of about 300 Gauss's magnitudes according to the maximum current of the 200mA that flows through little coil.

If the magnetic bead that interested sample comprises the magnetic susceptibility χ that is coupled to available traditionally, as to have about 4-5 μ m diameter and about 0.25 (for example, Dynabead) cell, then according to above formula (1), act on power F on the sample by about 300 Gausses' shown in Figure 9 peak field and be about 1 magnitude of receiving newton (nN).This power is enough to handle effectively such sample that adheres to magnetic bead.In other words, according to such power, the maximum fluid velocity that the sample of collection can bear is about the magnitude of 1 cel.In addition, under 37 ℃ biological compatible temperature (T=310K), the magnetic potential that is generated by the little coil with 200mA electric current can be the heat energy about 3 * 10 of such sample that adheres to magnetic bead ⁶Magnitude has doubly shown the strong capacity gauge of little coil.

Though above-mentioned example is based on the maximum current of the example that flows through little coil, but be to be understood that, much smaller electric current (for example, the magnitude of about 20mA) still provides enough peak fields and cause the power (for example, about 10 skin newton's magnitude) that is used for handling effectively various magnetic samples.Usually, the size of the magnetic force that is generated by little coil increases with the electric current that flows through little coil.In some cases, when the electric current increase was tending towards maximum current, the high current density on the period that prolongs in little coil can cause electromigration, and this is that wherein the big electric current in narrow conductor causes a kind of phenomenon of metal space (metal void) fault gradually.Though electromigration is more significant under higher temperature usually.Therefore, here (wherein working temperature typically is lower than 50 ℃ in the commingled system of Miao Shuing, and be adjusted to 37 ℃ for bio-compatibility in some cases), the current density that generates the magnetic force that is enough to be used in effective sample manipulation can not cause significant electromigration usually.

In addition, though above-mentioned example illustrates, be similar to those the little coils shown in Fig. 7 and 8 and can be provided for the tangible magnetic force that sample is handled, some concrete application may need even greater than above-mentioned magnetic force.Therefore, in another embodiment, permalloy (Permalloy), a kind of nickel alloy known traditionally, that comprise about 20% iron and 80% nickel can easily be magnetized and demagnetization, depend on that this alloy can be utilized around the electric current of its enhancing magnetic force in little coil design.Especially, in an exemplary manufacturing process, permalloy can (that is, have submicron-scale resolution) by using photoetching technique or electron beam lithography technology suitably to be deposited (for example, electroplating) on the little loop construction of multilayer.

According to another embodiment, " vertical " little coil can be made and can be used in and be handled and the magnetized sample of imaging, is similar to the little coil of above-mentioned multilayer.Current available CMOS technology is mainly supported the planar metal layer, therefore little coil discussed above is actually " flat ", because they are set up along the plane that is parallel to the x-y axle of representing on various figure, and generate magnetic field (that is, basically along the z axle) perpendicular to the surface of IC chip 102.Yet, in another embodiment,, plane micro coil might be tilted leave substrate surface (after removing oxide) by utilizing micromachined and/or other three-dimensional assembling process as the back making step, produce vertical little coil.Vertical little coil like this produces the magnetic field on the surface (that is, basically on the plane that is parallel to the x-y axle) that is parallel to IC chip 102.By utilize vertical and plane micro coil in according to an embodiment of present disclosure, it is possible that three-dimensional samples is handled, and also comprises rotation except straight line is carried.In the RF detection that discusses in more detail below and the context of imaging, vertical little coil can allow to be used for the RF disturbance of the large-signal of imaging, and plane micro coil provides dc fields to handle sample, strengthens the ability of the commingled system that has merged vertical and plane micro coil thus.

Discuss based on traditional semiconductor fabrication process, aspect the structure of exemplary little coil of present disclosure and making various after, be used to handle sample about generating magnetic field now, consider in more detail between the adjacent little coil in an array interaction.As discussed above, the principle of work of Fig. 6 (a) and the micro-coil array 200B that (b) illustrates is to create and move one or more magnetic fields peak value by the electric current in corresponding little coil 212 of modulation array, so that move and/or collect the magnetic sample.The size in the magnetic field that is generated by given little coil of array is based on the size of the electric current that flows through little coil, and each little coil can both be created on the local magnetic field peak value above little coil in array.Under this meaning, array 200B can be considered to " magnetic pixel " usually, and wherein the micro-coil array of N * N can produce N * N magnetic field peak value at least, or " pixel ", and each can both attract and collect sample.Figure 10 conceptually illustrates two adjacent little coil 212-1 and 212-2 of array 200B, and wherein the electric current 230 that equates basically flows through little coil, to be created on two magnetic field peak value 232-1 and 232-2 that equate basically on the coil.In Figure 10, the distance between two magnetic field peak values is usually corresponding to the spacing 216 of array 200B, as expression on Fig. 6 (a) and 10.

In one embodiment, the size that not only flows through the electric current of each little coil can be modulated so that the sample manipulation, and the sense of current that flows through given coil can also change, so that individual element ground carries out more level and smooth transfer to sample, or improve the spatial resolution (that is, reducing the spacing 216 of array effectively) that sample is handled effectively.Figure 11 (a)-(e) illustrates for adjacent little coil 212-1 of Figure 10 and five kinds of example scenario of 212-2, has the size of current and the direction of variation and generate the magnetic field that finally obtains in corresponding coil.Figure 12 illustrates every kind of situation for five kinds of example scenario shown in Figure 11 (a)-(e), the size of current in each coil and the figure of direction.On the transverse axis of Figure 12, stage 1-5 is respectively corresponding to five kinds of situations shown in Figure 11 (a)-(e).Top curve representation on Figure 12 flows through the electric current 230-1 of " left side " little coil 212-1 under every kind of situation, following curve representation flows through the electric current 230-2 of " the right " little coil 212-2 under every kind of situation.

Especially, in Figure 11 (a), as stage 1 expression of the curve map of Figure 12, the little coil 212-1 in the left side does not flow through its electric current, and the little coil 212-2 in the right has-and 20mA flows through its electric current.As a result, little on the right coil 212-2 goes up and generates magnetic field peak value 232-2.In an exemplary based on above little loop construction of discussing in conjunction with Fig. 7-9, the size of the magnetic field peak value 232-2 of Sheng Chenging can be about 30 Gausses' a magnitude like this.In Figure 11 (b), as stages 2 expression of Figure 12, the electric current 230-1 of the little coil in the left side is increased to about 12-13mA, and the electric current 230-2 of the little coil in the right is reduced to pact-19mA.Shown in Figure 11 (b), magnetic field begins broadening on two little coils a little, because some contributions from the little coil in the left side and the right are arranged now.

In Figure 11 (c), as stages 3 expression of the curve map of Figure 12, the little line in the left side and the right has the equal sizes that flows through them but the opposite electric current (about 17-18mA) of direction; As a result, generate wide peak, magnetic field, its center is the mid point between the center of corresponding coil roughly.In Figure 11 (d), the electric current 230-1 among the little coil 212-1 in the left side further increases, and the electric current 230-2 among the little coil 212-2 in the right further reduces, and at Figure 11 (e), electric current 230-1 is increased to 20mA at last, and electric current 230-2 is reduced to zero at last; As a result, the little coil 212-1 of on the left side goes up and keeps single magnetic field peak value 232-1.Should be appreciated that each magnetic field equal and opposite in direction that on Figure 11 (a) and 11 (e), is generated but direction is opposite.Therefore, by changing the electric current of the different directions flow through coil gradually, the magnetic field peak value can move between two adjacent coils continuously, therefore strengthens the resolution of array effectively, is convenient to the level and smooth transfer on array 200B of accurate localization and sample.

Discuss in conjunction with Fig. 1 and 2 as above, in one embodiment, various control assemblies 400 that are used for controlling and be dispensed to the electric current (and/or voltage) on little coil of array 200B can merge with the array of IC chip 102.In one embodiment, these control assemblies comprise one or more current sources (and/or voltage source) and various exchange or multiple connection parts, (with computer programmable) control so that the numeral of the field that is generated by array 200B.

Figure 13 is the micro-coil array 200B and various the control assemblies related with this array 200B that illustrates according to an embodiment of present disclosure.In the example of Figure 13, array 200B comprises " little coil unit " 250 of 8 row and 8 row, and wherein each little coil unit comprises little coil 212, and switch and logical circuit, as what further discuss below in conjunction with Figure 14 and 15.For electric current (and/or voltage) is distributed on little coil unit 250, the array 200B of present embodiment is divided into four quadrant 200B-1,200B-2,200B-3 and 200B-4, each quadrant has 16 little coil units 250 (that is, each quadrant 4 row and 4 row).Yet, should be appreciated that the micro-coil array according to present disclosure is not limited to this aspect with relevant control assembly, and concrete configuration shown in Figure 13 mainly is provided to be used for explanation.

As shown in figure 13, various related with array 200B in the present embodiment control assemblies comprise: line decoder 460-1, and it provides enforcement energy signal (row enable signal) R0-R7 to give the corresponding line of array 200B; Column decoder 460-2, it provides row enable signal C0-C7 respective column to array.Line decoder receives with binary-coded three digital row selection signals 466 (Row Select[0:2]) as input, exercises energy signal R0-R7 so that generate of wanting in any given time.Similarly, column decoder receives with binary-coded three digital array selecting signals 464 (Column Select[0:2]) as input, so that a row enable signal C0-C7 who wants in any given time generation.Line decoder 460-1 and column decoder 460-2 receive common clock signal 462 (Clk), be used for given enforcement can signal and the generation of given row enable signal synchronous so that at a specific little coil unit 250 of given selection of time.In an exemplary, clock signal 462, row selection signal 466 and array selecting signal 464 are provided by one or more processors 600, discuss in conjunction with Fig. 1 and 2 as above, like this, these signals can be generated according to computer control able to programme and/or that the user selects.

Figure 13 also conceptually illustrates four variable current source 420-1,420-2,420-3 and 420-4, and they provide the little coil unit 250 of controllable variable current to array 200B.A current source of the example of these four current sources, i.e. variable current source 420-1 is shown as and is configured to receive three digital current level signal 468-1 (Current Level[0:2]) and control voltage 469 (V _CTRL), and provide controllable variable current 470-1 (I1) as output to array.As what further discuss below in conjunction with Figure 16, in one embodiment, variable current source 420-1 is configured to according to the current level signal 468-1 of digital binary coding and control voltage V _CTRLVoltage one of eight different electric currents are provided.In the configuration of Figure 13, though not significantly expression on the figure, other current source 420-2,420-3 and each current source of 420-4 also receive three binary-coded digit current level signals and control voltage V _CTRLAs input, and provide and have eight different possible current levels, corresponding variable current output.Aspect of present embodiment, the digital current level signal of each variable current source can be provided by one or more processors 600, discuss in conjunction with Fig. 1 and 2 as above, like this, these signals can be generated according to computer control able to programme and/or that the user selects.

At last, Figure 13 also illustrates, the array 200B of present embodiment receives for the common direct supply voltage Vdd of all little coil units 250 of array, and to receive for all little coil units 250 also be " direction " signal 472 (Dir) of the sense of current (polarity) of common, as to have determined to flow through each little coil unit 250 little coil.This direction signal 472 is further discussed in conjunction with Figure 14 and 15 below.

Aspect of the embodiment of Figure 13, variable current source is configured to make each current source to provide electric current to all little coils in the quadrant of array with respect to little coil unit.For example, in one embodiment, current source 420-1 provides the little coil of electric current to first quartile 200B-1, and current source 420-2 provides current to the second quadrant 200B-2, current source 420-3 provides current to third quadrant 200B-3, and current source 420-4 provides current to four-quadrant 200B-4.In this configuration, each quadrant of array 200B is worked in substantially the same mode; Therefore, discuss a quadrant of array now in more detail.

Figure 14 illustrate according to an embodiment of present disclosure, parts is various interconnected in the first quartile 200B-1 of array 200B shown in Figure 13.The enforcement that is provided by the line decoder 460-1 of Figure 13 can signal R0-R3 be displayed on the left side of Figure 14, and the row enable signal C0-C3 that is provided by the column decoder 460-2 of Figure 13 is displayed on the top of Figure 14.First quartile 200B-1 comprises and is arranged in 4 row and 4 row and is coupled to 16 identical little coil units 250 on enforcement energy signal and the row enable signal.Each little coil unit 250 also is coupled to direction signal 472 (this signal is shared by the All Quardrants of array), and variable current source 420-1, and it provides controlled variable current 470-1 (I ₁) to all little coil units of quadrant 200B-1.Just also as shown in figure 14, each little coil unit 250 comprises logical and (AND) door 460-3, it the enforcement corresponding with the unit can signal and the row enable signal coil enable signal 474 is provided when all existing.Coil enable signal 474 is added to little coil crosspoint 460-4, and it comprises little coil 212 and be used for controlling the various switches of the electric current that flows through little coil after applying coil enable signal 474.

Figure 15 illustrates the content of little coil crosspoint 464-4 shown in Figure 14.Each little coil crosspoint comprises little coil 212 of being connected to direction of current (polarity) switch 460-5 (S1) and coil enable switch 460-6 (S2) (for example, being similar to above those that discuss in conjunction with Fig. 7-12).Supply voltage Vdd is added to polarity switch S1, and provides variable current source (being represented as C in Figure 15) to the connection of coil enable switch S2, allows electric current 470-1 to flow through coil with convenient switch S 2 when closed.Polarity switch S1 is by direction signal 472 controls, and coil enable switch S2 is by 474 controls of coil enable signal; Especially, when can signal corresponding to the enforcement of the little coil unit that comprises little coil and row enable signal when all existing, coil enable signal 474 makes and switch S 2 closures flows through little coil 212 to allow electric current 470-1.Aspect of present embodiment, direction signal 472 can be provided by one or more processors 600, discusses in conjunction with Fig. 1 and 2 as above, and like this, this signal can be generated according to computer control able to programme and/or that the user selects.

Figure 16 illustrates provides controllable variable current 470-1 details to the variable current source 420-1 of the first quartile 200B-1 of array.Again, on Figure 13, other current source 420-2,420-3 and 420-4 can implement in the same manner with current source 420-1.According to an embodiment, current source 420-1 comprises current level code translator 422-1, it receives digital binary code current level signal 468-1 and provides 8 to enable output, with the switch (in an exemplary, current level code translator 422-1 can utilize " thermometer-code (thermometer code) ") of selectively closed 8 switch 424-1A in the 424-1H.One side of each switch is connected to " substantially " current source, like this, 8 different baseline current-source 426-1A is arranged to 426-1H.Each switch 424-1A jointly is connected to the opposite side of 424-1H, so that controllable variable current 470-1 (I to be provided ₁), this variable current has one of 8 different possible current levels (that is I, in any given time ₁Be certain multiple of the electric current that provides by given baseline current-source).

Aspect of present embodiment, each baseline current-source 426-1A can be implemented by using MOS transistor in a conventional manner to 426-1H, and wherein the electric current that is provided by each basic source is by control voltage 469 (V _CTRL) determine.For example, in an exemplary, control voltage V _CTRLCan be added to all baseline current-source, like this, specific control voltage provides corresponding electric current from each basic source (for example, 0.7 to 3.3 volt control voltage generates the corresponding electric current each basic source of from 0 to 1.3 milliampere).Should be appreciated that in different embodiments control signal V _CTRLCan be changed,, or alternatively, can be retained as constant (for example, being connected to Vdd) so that variable fundamental current is provided.

In addition, be configured to provide 8 different current levels, be not limited thereto at present disclosure aspect this though should be appreciated that variable current source 420-1 shown in Figure 16; Promptly, be similar to the sort of general configuration shown in Figure 16 and can be implemented the current level that comes to provide according to a plurality of baseline current-source different numerical value, this numerical value can be via being similar to code translator shown in Figure 16 according to digital signal by having suitable bit number of the number of the current level that will be provided but selectable.In another embodiment, pulse width modulating technology can be utilized to by using single baseline current-source that variable current 470-1 is provided.In such embodiments, the fixed current that is provided by single source is become to have different duty factors by pulse-length modulation, the wherein relatively low lower average current of duty factor representative, and the higher average current of higher relatively duty factor representative.In one aspect, the numerical value that is used to provide the possible duty factor of different average current level can be determined to be similar to the sort of mode that adopts in the configuration of Figure 16, the digital binary coded signal that wherein is added to code translator provides a plurality of differences possible duty factor, thereby different electric currents is provided.

In the above embodiment that discusses in conjunction with Figure 13-16, comprise that various the control assemblies of variable current source, exchange and multiple connection parts, logic gate or the like are utilized as " digital switching network " of controlling and distribute the electric current among the micro-coil array 200B effectively.Aspect of these embodiment, such digital switching network makes that the control of array 200B is more feasible, particularly number (the N of little therein coil unit 250 ²) may be in sizable embodiment; More particularly, electric current can the mode with multiple connection be shared between a plurality of little coils in time, and the input of the digital signal of less relatively number can be utilized to control whole micro-coil array.Referring again to Figure 13, needed in the present embodiment, array control is provided and is convenient to the signal that sample handles and comprise 462,12 current level signals of 464,3 row selection signals of 462,3 array selecting signals of clock signal (promptly, 3 signals that are used for each current source of four variable current sources are as what represented by the signal 468-1 that is used for one of current source), be used for the control voltage 469 (V of current source _CTRL) and direction (polarity) signal 472.As discussed above, any one or all above-mentioned signals can be provided by one or more processors 600, and as shown in above Fig. 1 and 2, like this, these signals can be generated according to computer control able to programme and/or that the user selects.

On Figure 13-16, various control signals are provided to usually make that a little coil of array can generate the magnetic field with the possible field intensity of difference in any given time according to the variable current that flows through little coil.Therefore, aspect of present embodiment, in order to generate a plurality of magnetic fields so that a plurality of samples are collected or moved to " side by side " effectively, compared with sample for having magnetic field or not existing on the faster time scale of " reaction time " in magnetic field, different little coil of array is sequentially enabled (that is the electric current multiple connection of near little coil).Like this, the magnetic field of order generation may appear to simultaneously for the sample of being discussed and generate.A plurality of coils of array can sequentially enabled (for example, under computer control) according to each agreement in various " scan protocols " on the reasonable time yardstick; For example, in an exemplary, traditional " raster scanning (raster scanning) " agreement can be utilized to line by line each little coil of array is enabled in proper order, from the upper left corner of array shown in Figure 13, proceed to the right along first row, then to second row or the like.

For be provided for that sample handles suitable sweep time yardstick some exemplary illustrations, under the aqueous water environment, consider to have about 4-5 μ m diameter, commercially available magnetic bead (for example Dynabead) is as the magnetic sample of representing on the market.Usually, the sample that floats on a liquid stands viscosity resistance when they are mobile in liquid; This viscosity resistance influences the speed (also therefore influence " response time " of sample) of sample for the external magnetic field reaction usually.For the magnetic sample that floats on a liquid, response time τ _CutoffBe given:

τ _cutoff≈O(μμ _o/χB ²)，(2)

Wherein μ is the dynamic viscosity of liquid.Therefore, (that is, the inverse of the response time of sample in the pulsed magnetic field of) frequency, pulsed magnetic field presents for sample and applies continuous basically average magnetic force if sample is exposed to " cutoff frequency " that have significantly greater than sample.Like this, current source can be between a plurality of little coil of array with suitable speed by multiple connection (that is, sequentially applying in time), so that the sample aspect from being discussed generates the magnetic force that looks continuous.The magnetic force that obtains from magnetic field was usually discussed in conjunction with formula (1) in the above.For under the magnetic field of about 30 Gauss's magnitudes, has Dynabead diameter, in water of about 5 μ m, response time τ _CutoffBe about 10 ^-2The magnitude of second.Have pulsed magnetic field (for example, greater than about 100Hz) greater than the frequency of the inverse of response time of sample by use, the power that finally obtains equals the product of duty factor and the power that is provided by formula (1).

In case sample attracted to local magnetic field, just must keep enough magnetic potentials can collect sample in the field.Especially, the sample that floats on a liquid motion disorderly owing to the random collision of sample and fluid molecule on every side, this is a kind of phenomenon that is called as Brownian movement.Such Brownian movement meeting causes the diffusion of sample; Because Brownian movement, sample can move with random velocity (for example, in the mode back and forth of bending) on random walk in any given time, left its position.As discussed above, kinetic energy and the temperature relevant with this motion are proportional (that is, 3/2kT).Therefore, in order to keep collecting, the average magnetic potential of the field of generation can abundant heat energy greater than sample.

In view of above-mentioned content, in case sample initially is collected based on pulsed magnetic field, as long as do not interrupt at the internal magnetic field in period that allows sample be diffused into significantly away from " collecting zone " on given little coil, sample just can keep being collected in the pulsed magnetic field.For the magnetic field upper limit τ of break period _OffBy τ _Off＜d ²/ D is approximate to be provided, and wherein d is the diameter of little coil, D be sample diffusion constant (from the definition of D, for given time t, the particle mean distance d=(Dt) that advances ^1/2).The diffusion constant D of sample (square metre per second) is provided by following formula:

D＝kT/3πηa(3)

Wherein η is that the viscosity (kg/m.s) and the α of liquid are the diameters of sample.Under the example scenario of being considered, the viscosities il of water is about 10 ^-3The diameter of kg/m.s and Dynabead sample is 5 μ m; Therefore, suppose the temperature T (being room temperature) of about 300K, be about 8.5 * 10 for the diffusion constant D of the Dynabead sample in the water ^-14m ²/ s.If supposing little coil diameter is 20 μ m, τ _OffShould be less than about 5000 seconds.From the viewpoint of reality, above-mentioned example explanation, 10,000Hz or higher frequency (that is τ, _Off＜10 ^-4Second) the multiple connection electric current that is added to little coil does not in fact allow because the tangible diffusion of the sample that Brownian movement causes; For τ break period _Off＜10 ^-4Second, the Dynabead diffusion of 5 μ m is about only 3 nanometers.

Usually, should be appreciated that the configuration of the current source shown in Figure 13-16 and little coil and above-mentioned multiple connection technology are provided as exemplary, also is possible according to other configuration of present disclosure.For example, in the configuration of replacing, array 200B (for example can be divided into either large or small subdivision again, 4 little coil units of each subdivision, rather than 16), wherein the variable current at different current level each subdivision, that have predetermined value is provided by a current source for this subdivision special use.Alternatively, in another embodiment, have only such current source to provide current to all little coil units of array 200B in (for example, multiple connection) mode that time of order shares.In another configuration, each little coil unit can be equipped with its variable current source, like this, need be between a plurality of little coils current source of multiple connection.Usually, by using a current source to provide current to the dc power that any embodiment that a plurality of little coils utilize the electric current shared mechanism has reduced system.

It should also be understood that, though the above exemplary concepts of discussing in conjunction with Figure 13-16 concentrates on little coil by driven with current sources, based on the general exchange of Figure 13-16 general introduction and multiple connection structure, being used for embodiment that field that sample handles generates the replacement of array also can be based on using the little coil that driven by voltage source or the electric field generation and the dielectrophoresis principle of microtrabeculae.

For example, at first consider the micro-coil array 200B and relevant control assembly of Figure 13, replace variable current source with one or more variable (or fixing) voltage source.In such exemplary, corresponding little coil unit 250 of array with the top identical mode of discussing in conjunction with Figure 13-15 selected/enable.Yet, not that to allow variable current flow through selecteed/little coil of enabling, but that variable voltage (having selectable polarity according to direction signal 472) can be connected to is selecteed/little coil of enabling, so that (for example generate corresponding electric field from little coil, variable voltage source can replace variable current source 420-1 shown in Figure 14, and select/enable given little coil and little coil polarity via signal 472 after, variable voltage in fact be added to little coil on supply voltage Vdd connect).One or more variable voltage sources of such replacement embodiment can be implemented by traditional configuration (for example, digital-analog convertor) of any number that is applicable to various integrated circuit fabrication processes.

In another example that generates based on electric field, the micro-coil array 200B of Figure 13 can be above in conjunction with Fig. 5 (a) and (b) the micro-pillar array replacement of such appropriate size of discussion by being similar to, again, variable current source is replaced by one or more variable (or fixing) voltage source.In another example, the micro-coil array 200B of Figure 13 can be utilized to provide subsystem in conjunction with variable current source and variable voltage source, and this subsystem can carry out sample according to electric field and magnetic field and handle.These and other type, can be used in based on electric field or based on the embodiment of electric field/magnetic field and to relate to the various application that merge microelectronics and manipulation, sensing and the imaging system of miniflow.

III. Pattern detection, imaging and sign

Discuss in I joint as above, referring again to Fig. 1, various embodiment according to present disclosure, the field control assembly 400 of the commingled system of based semiconductor/miniflow also can comprise radio frequency (RF) and other detection part 480 that is coupled between generating unit part 200 on the scene and the one or more processor 600, is used for being convenient to being comprised in detection, imaging and sign one of sample of micro-fluidic system 300 or multinomial.Aspect different, RF/ detection part 480 based on higher relatively frequency (for example is configured to be convenient to, RF, microwave) electric signal (voltage or electric current) generates parts and generates electromagnetic field from the field, and is convenient to meter and is shown in the RF field of generation and certain type interactional signal between one or more interested sample.

Usually, as known according to Maxwell equation, but the RF field energy enough almost with under the RF signal frequency, conduct electricity or at any (biological or other) particle interaction of electric or magnetic polarization.Therefore, in the various embodiment of present disclosure, can singly discussing in the II joint, not be utilized to mobile sample in the interaction between RF electric field and/or magnetic field and the interested sample, but also can determine the position (for example, being convenient to imaging) of sample as above.

For example, the conduction sample has the ring current by the induction of RF field, and it produces their magnetic field again, and interacts consumingly with the field that adds.The existence that this interaction can be used for mobile sample and detect them.In the mechanism that discusses in more detail below, (for example, little coil of array) inductance, and this inductance value changes again can be by using high-frequency signal detected near the coil of the magnetic polarization of sample change sample.In another example, the electric polarization of sample causes the power that causes dielectrophoresis (DEP).This polarization can be via the change of the capacitance between the electrode (for example, having microtrabeculae or little coil of the voltage that applies) of sample and electric field generating apparatus, does not have the ground of dissipation, or by because the change of the decay that the electric polarization of vibrating in the sample causes is detected.

Above-mentioned example provides the various mechanism that can be used for detecting sample position.Based on detecting sample generates position component with respect to given field ability, in one embodiment, each generates the imaging pixel that parts 200 are similar to provides the important information that is used for making up the synthetic image that is suspended in the sample distribution in the micro-fluidic system (for example, considering the two-dimensional CCD array).In another embodiment, the image of sample distribution can be used as feedback again, handles one or more samples with algorithm according to the rules.

In an embodiment based on the sample that adheres to magnetic bead, the magnetic of magnetic bead is utilized to be convenient to pattern detection to the influence of the inductance value of little coil.For example, the inductance value L of given little coil and effective permeability μ _EffProportional.Near little coil during without any magnetic particle, μ _EffEqual space permeability μ ₀, but have certain magnetic permeability mu in existence _BeadMagnetic bead (for example, paramagnetic particles or PMP) time, the effective permeability related with little coil is μ _Eff=(1-α) μ ₀+ α μ _Bead(wherein α＜＜1), the inductance value with little coil changes certain amount Δ L thus.Therefore, monitor the inductance value L of little coil by being applied to high-frequency signal on little coil, such change Δ L of the inductance value of little coil can be detected, indicates thus to have the sample that adheres to magnetic bead near little coil.

Thereby depend on size and depend on the inductance value of little coil and the magnetoconductivity of magnetic bead, the scope of the change Δ L of inductance value can from L about 0.1% to L 1% (for example, have about 4.5 to 5 microns diameter and about 1.25 μ ₀Magnetic permeability mu _BeadMagnetic bead Dynabead can cause the change Δ L of inductance value of about 0.1% magnitude of L).In addition, also should consider the frequency response of magnetic bead magnetoconductivity; Especially, for Dynabead example, μ _BeadHas real number value for the frequency that is lower than about 100MHz.Therefore, in an exemplary, be lower than or the RF signal that approaches 100MHz is used in detection scheme.

Figure 17 is a setting embodiment, that be used to be convenient to RF/ detection part 480 pattern detection, formation " frequency-locked loop " that illustrates according to present disclosure.In the embodiment of Figure 17, exemplary little coil 212 is by its variable inductance L (it changes when having the magnetic sample) and its relevant coil resistance R _LBe expressed.Aspect of present embodiment, variable inductance L and coil resistance R _LForm the part of bridge diagram 485, this bridge diagram also comprises known predetermined capacitor C _RF(has dead resistance R _C) and two known resistance R ₁And R ₂

In order to be easy to illustrate and for the ease of following discussion, the remaining part among Figure 17 is shown as and is directly connected to little coil 212; Yet, be to be understood that, in other embodiments, remaining RF/ detection part 480 shown in Figure 17 can be shared together with the mode of other circuit that DC current is provided to little coil with multiple connection between a plurality of little coil of micro-coil array, is used for sample as discussed above and handles.For example, can use another signal that is similar to direction signal 472, if (suitable) (for example selects signal and extra switch together with row and column, to be similar to as above mode discussing in conjunction with Figure 13-15) so that use the RF of multiple connection and DC signal to be used to handle little coil manipulation with testing goal.

As mentioned above, in the embodiment of Figure 17, " frequency-locked loop " is made up of bridge diagram 485, phase detector 482, low-pass filter 484 and voltage controlled oscillator (VCO) 486.In general, phase detector, low-pass filter and voltage controlled oscillator are similar to the known parts that find traditionally in phase-locked-loop configuration.Yet, combination together with the parts shown in other, causes the lock-in circuit based on frequency rather than phase place to comprise the bridge diagram that is provided with uniquely of little coil 212 together, wherein the change direct correlation of locking frequency and inductance L ground changes, and wherein inductance L is owing to the existence of sample changes.Therefore, the change of the locking frequency by monitoring circuit shown in Figure 17 can detect near the existence of sample little coil.

For the work of circuit shown in Figure 17 is described, we at first consider an exemplary, and wherein the output V (ω) of VCO 486 is as the control voltage V that is input to VCO _cThe voltage of sinusoidal variations function, that have angular frequency.By using phase place complex vector symbol Ae ^{J θ}Represent sinusoidal voltage (wherein on behalf of amplitude and θ, A represent phase place), and expression is all about the phase place of the output of VCO486, the voltage V that obtains from bridge diagram 485 ₁(ω) and V ₂(ω) can be represented as V ₁e ^{J θ 1}And V ₂e ^{J θ 2}, wherein

θ 1 = - \arctan \frac{ωL}{R} - - - (4)

And

θ 2 = \arctan \frac{1}{ω C_{RF} (R_{2} + R_{C})} . - - - (5)

As what discuss in more detail below, frequency-locked loop is configured to make controls voltage V when θ 1=θ 2 _cBe stabilized in certain DC value.Therefore, from above formula (4) and (5), be used for " locking frequency " ω of frequency-locked loop _LockCan be represented as:

ω_{lock} = \sqrt{(\frac{1}{L C_{RF}}) (\frac{R_{1} + R_{L}}{R})} . - - - (6)

From foregoing, be appreciated that locking frequency ω _LockCome down to the function of the variation of little coil inductance L, because C _RF, R _L, R _C, R ₁And R ₂It is known fixed numbers.In an exemplary, consider the little coil inductance L of nominal of about 1nH magnitude, and the nominal coil resistance R of about 50 Ω _LIn order to ensure ω _LockBe lower than or approach 100MHz, C _RFBe selected as 1pF, and the typical R of about 1k Ω magnitude _C, R ₁Be selected as about 50 Ω and R ₂Be selected as about 10k Ω.

The inductance that causes for the existence of measuring owing near magnetic sample little coil changes Δ L, measures the Snap lock frequencies omega _LockAnd it is compared with representing the non-existent nominal locking frequency of magnetic sample.ω therein _LockNominal ground is not about in the exemplary of 100MHz when the magnetic sample does not exist, because the change Δ ω of the locking frequency that causes in the existence of magnetic sample _LockCan be to be about 50 to 100kHz magnitude.In Figure 17, buffer amplifier 488 is utilized to a V (ω) and is transformed into square wave, can utilize edge counter 490 (for example, a series of triggers) to determine the change of frequencies omega for this reason.Especially, in one embodiment, edge counter 490 can be configured to the square wave during the given time interval along counting, and the numeral output of the such count value of representative is provided to the one or more processors 600 shown in Fig. 1 and 2, can determine the change of the frequencies omega that representative sample exists thus.

In circuit shown in Figure 17 was provided with, the function of phase detector 482 was electric current I=K of output _θ(θ 2-θ 1), wherein K _θIt is certain constant.This electric current I is added to low-pass filter 484, and this wave filter has transfer function in the Laplace territory

Z (s) = \frac{s + z}{s (s + p)}, - - - (7)

Wherein z is zero point of transfer function and p is a limit.From above content, can see that because the existence of capacitor 484A, transfer function Z (s) is included in the limit of s=0 in the denominator.In the Laplace territory, can be given at the expression formula of controlling voltage Vc:

V _C(s)＝IZ(s)＝K _θ(θ2-θ1)Z(s).(8)

From above content, can see that at steady state (SS) (s=0), it is infinitely great that Z (s) is tending towards, thus, in order to ensure stable control voltage Vc, value (θ 2-θ 1) must go to zero under steady state (SS).Therefore, the capacitor 484A in low-pass filter 484 guarantees in fact, and when θ 1=θ 2, frequency-locked loop is stable, provide thus above provide at ω _LockExpression formula.

Figure 18 illustrates the further details according to phase detector 482 embodiment of present disclosure, in the frequency-locked loop shown in Figure 17.As shown in figure 18, phase detector comprises two phase comparators 4821 and 4822, respectively is designed according to exporting " adding (up) " signal or " subtracting (down) " signal in the phase relation that is added between two signals of comparer.For example, the number of winning the confidence V (ω) leads over reference signal as being applied to the reference signal of each comparer if be added to another input signal of comparer, " adding " signal of then given comparer output pulse width modulation; Alternatively, if another input signal lags behind reference signal, " subtracting " signal of comparer output pulse width modulation then.Add with the duty factor of cut signal proportional accordingly with the amount that leads or lag accordingly.

According to the configuration of bridge diagram shown in Figure 17 485, can observe, in phase detector shown in Figure 180 482, signal V ₂(ω) always lead over phase theta 2 of reference signal V (ω), and signal V ₁(ω) always lag behind phase theta 1 of reference signal V (ω).Therefore, in embodiment shown in Figure 180, " adding " signal of phase comparator 4821 never activates, and therefore remains not to be connected in the circuit; Similarly, " subtracting " signal of phase comparator 4822 never activates, and therefore remains not to be connected in the circuit.Figure 19 is illustrated in the further details (phase comparator 4822 is configured to be similar to comparer 4821) of the phase comparator 4821 of phase detector shown in Figure 180 482.As shown in figure 19, phase comparator 4821 comprises two d type flip flops and is coupling in the corresponding Q output terminal of trigger and the logical AND gate (AND gate) between the RESET input (R).

Aspect of the embodiment of Figure 18, from the plus signal of comparer 4822 according at V ₂(ω) and the activating transistor 4824 periodically of the phase-lead amount between the V (ω), to allow by current source 4823 electric current I that rises; Like this, referring again to Figure 17, the capacitor of low-pass filter 484 basis is at V ₂(ω) and the phase-lead amount between the V (ω) " pumped into " with electric current.Similarly, from the cut signal of comparer 4821 based at V ₁(ω) and the activating transistor 4825 periodically of the phase lag amount between the V (ω), with based at V ₁(ω) and the phase lag amount between the V (ω) extract electric current (to ground) from the capacitor of low-pass filter.In steady state (SS), the combined moving that pumps into and extract of electric current causes net current I to equal zero, corresponding to condition θ 2=θ 1.

Figure 20 illustrates the setting according to a kind of replacement of RF/ detection part 480A another embodiment of present disclosure, that be used to be convenient to pattern detection.A kind of homodyne detection of the representative system that is provided with of Figure 20, wherein two of frequency synthesizer 4802 voltage controlled oscillator VCO (I) and VCO (Q) generate sin ω t (homophase, or I) and cos ω t (quadrature mutually, or Q) signal respectively.The little coil 212 of homophase (sin) RF signal excitation, it changes the phase place and the amplitude of pumping signal then.The response of little coil (output of low noise amplifier, or LNA) be multiply by the original in-phase signal in the frequency mixer 1 then and is multiplied by quadrature-phase in the frequency mixer 2.The direct current output (OUT 1) of frequency mixer 1 and the dead resistance R of little coil _LProportional, and the direct current of frequency mixer 2 output (OUT 2) is proportional with the inductance L of little coil.Therefore,, can determine because the change of little coil inductance that magnetic sample (for example, adhering to the cell of magnetic bead) causes, indicate the existence of sample thus by monitoring OUT 2.

Aspect of embodiment shown in Figure 20, suppose as discussed above, under some example scenario, can hang down 0.1-1% because the inductance of little coil of causing of single magnetic bead changes Δ L, Design of Low Noise is for realizing that high precision RF sample sensing can be very favourable.Therefore, in an exemplary, frequency synthesizer 4802 can be implemented based on the very low noise high frequency oscillator of coplanar striplines (coplanar striplines) by using, be similar in July in 2004 19 and submit to, exercise question is the non-temporary patent application No.10/894 of the U.S. of " Methods and Apparatus Based onCoplanar Striplines ", submit to 674 and 2004 year July 19, exercise question is the non-temporary patent application No.10/894 of the U.S. of " Methods and Apparatus Based on CoplanarStriplines ", those that discuss in 717.

After the detection that the magnetic sample is discussed, each conception of species of disclosed herein, relevant RF field similarly can be used in discerns and characterizes interested sample.For example, by using the knowledge from the behavior of various materials in electromagnetic field of traditional solid state physics, the change that depends on frequency of the electric polarization of sample or magnetic polarization can be used for the type of recognition sample.These changes can be characterized on very wide frequency range.Therefore, in one embodiment, add the RF frequency (or use more exquisite signal processing technology) of the signal on the generating unit part of showing up by scanning, the frequency response of sample (for example, absorption spectra) can be measured in certain location, and can or characterize sample based on the response identification that measures.

Relevant RF field apply and under the control of RF/ detection part 480 among interactional another embodiment of sense field/sample, the RF field can be used for local measurement and be applied to magnetic resonance in the uniform magnetic field on the sample.Especially, the spin or the magnetic domain of given sample vibrate with characteristic frequency, and this can be used for discerning the type of spin type or sample itself.The magnetic resonance type comprises ferromagnetic resonance (FMR) (little YIG ball can be used as magnetic bead, and each YIG ball has single magnetic domain, and it rotates freely, because magnetic bead is spherical) on the GHz frequency.In addition, electron spin resonance (ESR) technology can be used for discerning and relate to the origin that characterizes them (that is the g factor of) spin, sample, and nuclear magnetic resonance (NMR) are used for discerning the g factor of nuclear spin.Therefore, according to principle discussed here, magnetic resonance imaging (MRI) system can be implemented on chip.

IV. Adjustment

Mention in conjunction with Fig. 1 and 2 as above, according to an embodiment, commingled system 100 can comprise temperature controlling unit 500.The field that is comprising big figure generate parts 200 very much and the exemplary of the field control assembly 400 followed in, the power consumption of system can be significantly, and the work of these parts can improve system in and system's temperature on every side.In view of above-mentioned content, the temperature of system can be adjusted to or near specific temperature, so that the bio-compatibility of system and cell/sample of being studied, and reduce risk foregoing, electromigration failures.

More particularly, according to an embodiment as shown in figure 21, temperature controlling unit 500 can comprise one or more monolithic temperature sensor 500A and the outer temperature controller 500B of sheet.At this moment referring again to Fig. 2, in different embodiments, a plurality of monolithic temperature sensor 500A can be set in the IC chip 102 and various position on every side; In Figure 21, an exemplary temperature sensor 500A usually is shown as in the environment that is in IC chip 102, and chip is coupled to enclosed chip 110 again.Aspect of present embodiment, one or more upper sensor 500A provide one or more temperature signal T _ChipTo processor 600, for the purpose of illustrating, processor is shown as comparer on Figure 21, with this signal T _ChipWith reference temperature signal T _RefCompare (in an exemplary, T _RefCan represent 37 ℃ temperature).

In various embodiments, processor 600 can be configured to receive a plurality of temperature signals from corresponding different sheet upper sensor, and (for example handle a plurality of signals according to one or more predetermined algorithm, on average, based on weighted mean of chip position or the like), so that the sensed temperature value of certain total to be provided, then it and reference temperature are compared.Comparative result based on one or more sensed temperature and reference temperature, control signal is provided to the outer temperature controller 500B of sheet, this controller correspondingly heats or (for example cools off enclosed chip 110, in an exemplary, thermoelectric (al) cooler or " TE " refrigeratory can be used as chip outer controller 500B).At present embodiment on the other hand, the heat conductivity with in every layer of passing all layers of IC chip 102 is such, and it makes can suppose that total system is in identical temperature.Therefore, regulation loop is that to be enough to keep the temperature of overall system be steady state value.

In the embodiment of Figure 21, exemplary built-in temperature sensor 500A comprises parasitic pnp bipolar transistor 5002 and reference current source 5004 (can obtain) in any standard CMOS process.If transistorized emitter current is held constant at reference current I _Ref, then transistorized emitter base voltage is given:

V_{EB} = - \log (\frac{I_{ref}}{I_{s}}) . \frac{KT}{q}, - - - (9)

Wherein the end of logarithm is e, I _sBe transistorized leakage current, k is the graceful constant of bohr thatch, and q is an electron charge, and T is an absolute temperature.Above formulate emitter voltage can be used as chip temperature (T _Chip) direct tolerance.In one embodiment, processor 600 by use 1 bit comparator with this emitter voltage with represent reference temperature (for example, T _Ref=37 ℃) the voltage of calibration compare.If T _Chip＞T _Ref, then the control signal operating temperature controller 500B that is provided by processor is so that cooled wafer, and vice versa.

In various embodiments, the precision of temperature regulator and long-time stability may be subjected to the mismatch of integrated component, the influences such as drift, I/f (flicker) noise and mechanical stress of parameters of operating part.In order to improve the precision of temperature regulator loop, in certain embodiments, can utilize various traditional Analogous Integrated Electronic Circuits designing techniques, such as automatic zero set (AZS), adaptive calibration and dynamic element coupling, and signal copped wave is with average.

V. Micro-fluidic system

Referring again to Fig. 1 and 2, in case comprise that of generating in parts 200, a control assembly 400 and the temperature controlling unit 500 or multinomial IC chip 102 are made, micro-fluidic system 300 just can be coupled on the IC chip 102 with formation commingled system 100.Discuss in I joint as above, according to an embodiment, micro-fluidic system 300 can be configured to better simply relatively chamber or liquid storage pool, is used to keep comprising the liquid of interested sample.For example, as what usually illustrate on Fig. 1 and 2, the miniflow liquid storage pool with substantial rectangular volume can comprise access pipeline 302 and 304, so that fluid flows into and the outflow liquid storage pool.Alternatively, micro-fluidic system can have setting more complicated, that comprise a plurality of pipelines or passage, and the liquid that comprises sample therein can flow, and various parts (for example, valve, mixer), is used for guiding and flows.In different embodiment, in case finish the semiconductor fabrication process, micro-fluidic system 300 just can be fabricated on the top of the IC chip 102 that comprises other system unit, to form commingled system 100; Alternatively, micro-fluidic system 300 can be made (for example, using the soft lithographic technology) discretely, is subsequently attached on the one or more IC chips that comprise other system unit, to form commingled system 100.

Others at embodiment shown in Figure 1, the electric field of commingled system 100 and/or magnetic field generate parts 200 and can be provided with various layouts with respect to micro-fluidic system 300, so that in the field that is generated and be comprised between the sample in (or flowing through) micro-fluidic system and interact.In various embodiments, a generation parts 200 can be set near the micro-fluidic system along one or more physical boundaries of micro-fluidic system, and layout interacts with respect to the field-sample of micro-fluidic system along one or more Spatial Dimensions with permission like this.Usually, according to each conception of species discussed here, under electric field that generates parts 200 generations by the field and/or the control of CALCULATION OF MAGNETIC FIELD INDUCED AROUND machine, interested sample can move through micro-fluidic system along any paths almost, be collected or be held at the certain location place, and be rotated in some cases.Like this, can be used for various operations based on for example by processor 600 available programmability and computer control and changed neatly at the topology of " the virtual microscale pipe system " of interested sample.This handles the extremely strong instrument that provides for the accurate cell in better simply and more exquisite relatively operation/object.

Usually, the top layer of CMOS chip comprises silicon nitride or polyimide passivation layer, its objective is to prevent to be penetrated in the chip such as the such chemical element of sodium.According to an embodiment, micro-fluidic system 300 also can further be produced on the top of CMOS chip passivation layer, and wherein micro-fluidic system comprises the polyimide sidewall of the micro-patterning with shape of wanting, so that form the passage of guiding sample, or " mini raceway groove ".Figure 22-26 illustrate according to an embodiment of present disclosure, making as the various treatment steps part of commingled system, that relate to during based on the micro-fluidic system of polyimide.

Especially, Figure 22 illustrates the part of the semiconductor chip 104 that comprises single chip 102.In one aspect, the part of the substrate 104 shown in Figure 22 bigger semiconductor wafer of having made a plurality of chips 102 from it is cut; In an exemplary, each chip 102 has the yardstick of about 2mm * 5mm magnitude, and wafer substrate can be cut into a plurality of parts of the yardstick with about 15mm * 25mm magnitude.

In case be cut, comprise that respectively the corresponding substrate sections 104 of single chip 102 can be by spin coating with polyimide, then by using traditional lithographic techniques patterning.Because CMOS chip list surface layer generally includes polyimide passivation layer, can make micro-patterning, adhere to the polyimide sidewall of the passivation layer of similar material well.Figure 23 is illustrated in the example of polyimide layer 310 at the top of substrate 104, and wherein polyimide layer comprises fluid passage 316 and two holes 320 by using traditional lithography patterning.In different exemplary, the coating and the patterning process that are used for polyimide layer can be configured to height from several microns to several thousand micrometer ranges and the width that forms the fluid passage according to the requirement of given application.

Make fluid passage 316 in polyimide layer 310 after, according to an embodiment, the surface of fluid passage can randomly apply (for example, spin coating) with thin dimethyl silicone polymer or PDMS layer.PDMS is a biocompatible material, and its surface can be functionalized, so that promote or the prevention cell adhesion.For example, aspect of present embodiment, the oxidized surface of using fiber adhesion albumen (FN) to handle the PDMS of polymerization makes can be influencedly with the extracellular matrix protein micro-patterning, so that cell adhesion and expansion.On the other hand, the surface of handling PDMS with Pluronic F127 can stop protein adsorption, therefore stops cell adhesion.These corresponding characteristics can so that according to an embodiment of present disclosure, the guiding biological specimen down to the microfluidic channel of cell sorter (following in the VI joint, further discuss) be used for according to another embodiment of present disclosure, organize cell guiding between erecting stage to aspect certain location (below further discuss) different in two-dimentional microscale at the VII joint.In various embodiments, PDMS can be spun onto the lip-deep micron thickness layer in fluid passage, does not handle or imaging and do not sacrifice sample.

As shown in figure 24, the cover plate 312 of suitable shaping (for example, cover glass) can be coupled to polyimide layer 310, forms miniflow chamber or passage.On the one hand, the surface of the cover plate that will combine with polyimide layer can be coated with negative photoresist or ultraviolet solidifiable epoxy resin 314 (for example, SU-8, can be from Microchem, Inc.of Newt on, Massachusetts buys) so that the sealing between cover plate and the polyimide layer (for example via with the curing of ultraviolet light) to assembly.Figure 25 illustrates the assembly of finishing of the cover plate 312 that is attached to polyimide layer 310, is used for enclosing fluid passage 316, and therefore forms micro-fluidic system 300.

At last, as shown in figure 26,

insert pipeline

302 and 304 and be coupled to the hole 320 of assembly, to finish micro-fluidic system 300 via pipe fitting 305.In one embodiment, the photoresist of UV-curable or epoxy resin can be used for a pipe fitting and pipeline once more and joins on the assembly.In Figure 26, the part of pipeline 304 is cut out on sectional view, flows with the fluid that shows the micro-fluidic system that passes through commingled system 100.

Figure 27-32 illustrate according to another embodiment of present disclosure, at the various treatment steps that comprise based on the manufacturing micro-fluidic system 300 of ultraviolet solidifiable epoxy resin patterning the time.In the present embodiment, at first with reference to Figure 27, each IC chip 102 (for example, having yardstick and about 270 microns thickness of about 2mm * 5mm magnitude) is adhered on the silicon chip 1040, and this silicon chip is different from the substrate that is used to make the IC chip.In other words, in the present embodiment, the IC chip is cut from bigger wafer, and these IC chips are made into the size (that is, not having extra substrate around chip area) that is substantially equal to their making floorages (footprint) on wafer on this wafer.Qie Ge chip is adhered on another bigger silicon chip 1040 (for example, having the yardstick of about 25mm * 25mm magnitude) then in this wise, and wherein bigger wafer can comprise the electrode that is fabricated on it so that with being electrically connected of IC chip.In one aspect, substrate 1040 can be used as enclosed chip 110 (see figure 2)s of commingled system.

As shown in figure 28, in the present embodiment, (for example, ground floor 318 SU-8), its thickness are thicker than the thickness (for example, about 300 microns) of IC chip 102 to the assembly of IC chip 102 and substrate 1040 a little with ultraviolet solidifiable epoxy resin by spin coating then.Via traditional optical lithography technology, a plurality of holes 320 are patterned in the ground floor, and the layer quilt baking (for example, 95 ℃ of following postexposure bakes 30 minutes) of patterning, but do not develop.Subsequently, as shown in figure 29, the second layer 322 of ultraviolet solidifiable epoxy resin is by spin coating (for example, to about 100 microns thickness), and is patterned via the optical lithography technology, to form the sidewall and the hole 320 of fluid passage 316.The same with ground floor 318, the second layer 322 is exposed the back baking, and the second layer is developed then, to form fluid passage 316.The development of the second layer makes the sectional hole patterns of ground floor 318 expose, and this hole liquid then is developed, to finish the formation in hole 320, as shown in figure 30.

Then, as shown in figure 31, glass or plastic cover sheet 312 coated thin layers with ultraviolet solidifiable epoxy resin (for example, 50 microns) cut into suitable shape, and are placed on the top of the assembly of patterning.The commingled system 100 of assembling (remove and insert pipeline) shown in figure 32, heated about 10 minutes down at 75 ℃, was applied to the epoxy resin on the cover plate 312 and was sealed in cover plate and the slit of the junction of second epoxy resin layer so that soften.Subsequently, the device of assembling is come blank exposure (blank-expose) with ultraviolet light, and is exposed back baking, to be solidificated in engaging between cover plate and the fluid passage sidewall.Then, insert pipeline to be connected on the assembly with the similar mode of the above Figure 26 of combination discussion.

According to another embodiment, commingled system 100 shown in Fig. 1 and 2 can implement that (details that is applicable to the soft lithographic technology of this embodiment is discussed: Younan Xia and George M.Whitesides are shown by using the liftoff making micro-fluidic system 300 of PDMS and soft lithography technical point and subsequently micro-fluidic system being attached to IC chip 102 in below with reference to document, at Annual Review of Material Science, 1998, Vol.28, " the Soft Lithography " that delivers on the pp.153-184, with George M.Whitesides, EmanueleOstuni, Shuichi Takayama, Xingyu Jiang and Donald E.Ingber are shown, at Annual Review of Biomedical Engineering, 2001, Vol.3, " the Soft Lithography in Biology andBiochemistry " that delivers on the pp.335-373).Figure 33-38 shows the various treatment steps that comprise when making micro-fluidic system based on such soft lithographic technology.

Shown in Figure 33 and 34, silicon chip with ultraviolet solidifiable epoxy resin 332, and is used photomask to be patterned via traditional optical lithography technology by spin coating, to produce fluid passage mould 334.In Figure 35, PDMS layer 336 is coated with (cast coat) to mould by casting, and by heat curing.As shown in figure 36, the PDMS layer of curing is peeled off from mould then, and the marking of fluid passage 316 forms therein.The PDMS layer is cut into the shape of wanting, and by drilling bore hole 320, to form micro-fluidic system 300.In Figure 37, be attached with the substrate 1040 coated thin layers 338 (for example, 50 to 100 nanometers) of IC chip 102 on it, to promote engaging between chip/substrate assembly and the PDMS micro-fluidic system 300 with silicon dioxide.Handle with oxygen plasma on the engaged surface of wanting of PDMS micro-fluidic system and chip/substrate assembly, so that the surface that " activation " is used to engage when exerting pressure, on Figure 38, the surface of activation is joined together, to form commingled system 100 (except the access pipeline).

In a word, according to various embodiment discussed above, the total manufacturing process that is used for CMOS/ miniflow commingled system can may further comprise the steps with suitable order according to employed concrete technology: comprise that 1) (for example, RF) casting of the silicon of the CMOS chip of electronic installation and interlock circuit and adjustment electronic installation is made for micro-coil array, digital switching network, imaging; 2) optional permalloy deposits on suitable little coil, with increasing magnetic field; 3) (for example, on chip, directly via based on polyimide or based on the technology of ultraviolet epoxy resin, or with soft rock version printing technology PDMS mold separation ground) makes micro-fluidic system; 4) PMDS with CMOS chip surface of various preparations for bio-compatibility applies; 5) apply cover plate, to form (a plurality of) fluid passage/chamber; And 6) with CMOS/ miniflow commingled system and electroplax (electricalboard) (for example, enclosed chip) and temperature controller (for example, thermoelectric (al) cooler) assembling.

VI. Sample counting and classification

According to another embodiment, as the above micro-fluidic system of in III joint, discussing that comprises pattern detection and image-forming block 100 and as the various configurations of the above micro-fluidic system of in the V joint, discussing can be used to various kinds of cell counting, classification and identification application.

For example, Figure 39 (a)-(d) illustrate via as the various exemplary of the cell detection of the above RF detection technology of discussing in conjunction with Figure 17-20.In 39 (a), single narrow microfluidic channel is once only to allow a sample that adheres to magnetic bead to cross given little coil (promptly, suspending liquid comprises the magnetic bead 112 that is adhered on the interested sample that flows through passage 300, this passage 300 is in the top that is coupled to little coil 212 of RF/ detection part 480 (coil 3), and this little coil is the sensing magnetic bead one by one).Cell count can be finished based on fluid flow rate that changes and the characteristic that is suspended in the magnetic bead in the fluid.In some cases, too fast if fluid flows, magnetic bead may not have time enough to magnetize in sensing coil (coil 3), therefore can not correctly be detected by RF/ detection part 480.Under such situation, can utilize other the little coil (for example, the coil 1 and 2 except coil 3) in the straight line micro-coil array shown in Figure 39 (a), generate D.C. magnetic field, to arrive sensing coil (coil 3) magnetized beads before at magnetic bead, be convenient to the detection of magnetic bead thus.

In another example, shown in Figure 39 (b), wide microfluidic channel 300 can be implemented, be coupled to above little coil 212 of RF/ detection part 480, this passage once transmits several magnetic beads 112, during this period, little coil 212 can be with a plurality of magnetic beads of count resolution while sensing of a magnetic bead.Other counting example provides on Figure 39 (c), wherein utilizes

fluid passage

300A, 300B and the 300C of multiple connection, and at the two-dimensional imaging that distributes via the magnetic bead of micro-coil array 200B and single liquid storage pool micro-fluidic system 300 shown in Figure 39 (d).As discussed above, by the RF/ detection part 480 of each little coil that can encourage two-dimensional array, micro-coil array 200B is similar to the pixel setting of traditional CCD imaging system.

According to another embodiment of present disclosure at based on accurate cytological classification method and apparatus according to CMOS/ miniflow commingled system various embodiment discussed above, that comprise the RF/ detection part.With the organ or tissue of high precision with even cell colony and decomposition, or keep apart with the blood that compiles in batch, for carrying out gene expression analysis, all being important for the cell of the pure cell line of needs and organizational project chemical examination or for clinical practice (stem cell that for example is used for the marrow-reconstitution process of cancer patient).Because the expression formula of unique cell surface receptor, many cells can be identified.In traditional method, coatedly be used for reversing the blood count art with magnetic bead and mesh (engage) cell with magnetic tweezer and magnetic at the ligand of these acceptors.This technology also is used in cytological classification/separation, but traditional simple fixed magnets of magnetic separation technique utilization, this magnet is once all added up the cell that classification one big group in ground is adhered to magnetic bead, shortage controllability and precision.Opposite with traditional method, an embodiment of present disclosure handles the microscale of the height controllability of CMOS electronic installation and micro-coil array and the detectability combination, with the cytological classification method and apparatus of realization for super accurate, high throughput that is attached to each biological cell on the magnetic bead in non-homogeneous suspending liquid and robotization.

In an exemplary of present embodiment, as shown in figure 40, the cell solution that comprises the cell 116 that adheres to magnetic bead and unmagnetized cell 114 (for example is suspended in the fluid that flows through microfluidic channel 300, in an exemplary, capillary endothelial cell and NIH 3T3 fibroblast can be suspended in have coated with in the medium at 2.8 microns magnetic bead of the antibody (a kind of cell surface receptor except endothelial cell) of PECAM (PECAM); Only be attached to the magnetic bead on the endothelial cell) coated with ligand.Microfluidic channel by RF sensor 212-1 or 212-2 (promptly, be coupled to the coil of RF/ detection part 480) top, the cell 116 that no matter when adheres to magnetic bead is during by the sensor top, sensor record and counting adhere to the cell of magnetic bead, aspect of embodiment shown in Figure 40, no matter when a RF sensor 212-1 detects when adhering to the cell 116 of magnetic bead, little coil in the first straight line micro-coil array 2000-1 all order activates, as " travelling belt ", to spur the cell 116 that adheres to magnetic bead, thus it is removed from the cell fluid stream of combination, and from total cell colony, isolate the cell that adheres to magnetic bead effectively.In one embodiment, straight line micro-coil array 2000-1 does not need always to be switched on, so that minimise power consumption, it can be switched on the signal of the cell 116 RF sensor 212-1 that exist, the front that represent to adhere to magnetic bead.

In Figure 40, in some cases, some cells 116 that adhere to magnetic bead can pass through the first straight line micro-coil array 2000-1, and can not be drawn out from the main flow of flow liquid.Yet, aspect of the embodiment of Figure 40, multisensor-straight line micro-coil array piece can sequentially be utilized, and each has identical operations agreement (for example, pointing out that little coil 212-2 is used as second " the RF sensor " and the second straight line micro-coil array 2000-1).Such redundant system with independent cell selection increases cytological classification output and precision substantially, and does not sacrifice speed.RF sensor 212-1 and 212-2 adhere to the existence of sample 116 of magnetic bead and real time monitoring nicety of grading quantitatively by sensing.Transmitting by after this system, cell that adheres to magnetic bead that separates and the cell that does not adhere to are collected respectively, the cell colony that does not wherein adhere to can be used for further classification by same agreement, to remove the cell may stay any in this colony and to adhere to magnetic bead (estimation may seldom).

The cytological classification method and apparatus of example explanation provides several important advantages compared with prior art in being provided with of Figure 40.For example, on the one hand, each cell that adheres to magnetic bead can be isolated from non-homogeneous cell colony with low-down error rate, and wherein precision quantitatively monitors in real time by using the RF/ detection part.And, in conjunction with the precision of the cytological classification method and apparatus of Figure 40 discussion compared with being developed and much higher by the clinical traditional magnetic separation technique that is used for specific blood cell type or pathogen and the blood that compiles are in batch kept apart (stem cell that for example is used for the marrow-reconstitution process of cancer patient) that makes.In traditional method, be filled pipe by use with the steel wool that surrounds by fixing magnet, the cell that a big group is adhered to magnetic bead once all statistics ground from remaining blood content, pull out.This method be labour intensity very high with lack precision, particularly when certain type cell needs " fully " when being eliminated.

In addition, cytological classification method and apparatus discussed above has been convenient to the microfluidic channel of multiple connection and the parallel fluid treatment of cmos circuit.The CMOS electronic installation also makes the cytological classification robotization become possibility.(FACS) compares with FACS, according to the system of notion discussed here can with much smaller and more inexpensive manner be implemented.And, need the minimum cell that is used to classify to prepare the transfection of fluorescence protein (for example, without) according to the cellular classification system of present disclosure.In addition, on the other hand, can prove with micro-fluidic system compared with the easier maintenance physiology of any bulky equipment homeostasis.

According to the various aspects of the embodiment shown in Figure 40, the consideration of a plurality of reality may influence the cytological classification process.For example, some variable that can influence cytological classification comprises, but not necessarily is limited to: the 1) efficient of ligand-receptors bind to the target cell; 2) occurrence rate of the non-specific binding of magnetic bead to the non-target cell; 3) number of cell type in the solution; 4) cell number of the density of Xuan Fuing or every liter; And 5) cell is from the tissue or the efficient that is decomposed of organ of results.First and second variablees can be solved for the specific ligand of the cell surface receptor of being represented uniquely by the target cell type by selection.

For example, when aiming at endothelial cell, PECAM is the desirable selection of cell surface molecule in an exemplary, because the expression of its uniqueness in endothelial cell and because its effect in cellular activity and cell adhesion; As a result, the possibility of the separation of during transfer accompanying magnetic bead reduces.In another embodiment, endothelial cell can be classified from also comprise the fibroblastic cell suspending liquid of NIH 3T3 that does not show PECAM.The through-rate of cell suspending liquid and density can be calibrated, so that reach the optimal classification performance.

In addition, in other embodiments, can utilize repetitive process, wherein in first assorting process optimised experiment parameter as the starting condition that is used for one or more later assorting processes, like this, cell can be classified from the suspending liquid that comprises the various kinds of cell type.For example, in involving a process of neonatal heart, endothelial cell can be separated with cardiac muscle cell, fibroblast, the immunocyte and the nerve fiber of overflowing before results.The environment of " noisy " that is caused by the cell colony of this mixing is determined the border of cytological classification performance in some cases.On the one hand, diluting cells suspending liquid can increase and is used to classify the needed time, but can improve nicety of grading.On the other hand, in order to ensure enough decomposition, suspending liquid can transmit by filtrator, and filtrator optionally filters the big cell mass of having avoided by trypsase and clostridiopetidase A dissolving.

VII. The tissue assembling

In another embodiment according to present disclosure, the assembling of the microscale of designed tissue can realize by using the whole bag of tricks discussed here and equipment.For example, in one embodiment, can finish by utilizing CMOS/ miniflow commingled system 100 discussed here from the designed heart tissue of heterocyst colony assembling microscale.

The media that the signaling dialogue of the complexity between the various kinds of cell type in space that reorganize by each developmental character step, closely restriction forms as tissue morphology.In the tissue of maturation, the space of these cell colonys and colony's control are kept to full capacity, but its loss is with the beginning of discernible mode sign lysis.Unknown is, seems how the trickle interaction of controlled cell colony can strengthen the morbidity incident.The fabulous example of this respect is the cell-cell interaction between kapillary and cardiac muscle fiber in the heart, and the potential propagation of its change activity causes influencing arrhythmia cordis.This is an important problem, because the ARR reliable measure of current drug therapy heart of no use clinically.And the antiarrhythmic drug streamline of medicine and biotech company is blank, and part is owing to lack the experimental sample of the identification of supporting new pharmaceutical target.Therefore, the ability that designs the microscale heart tissue of non-homogeneous cell colony provide the cardiac arrhythmia that is used to find new pharmaceutical target reliably, effective sample, and illustrated answer for the underlying issue in the cardiac electrophysiology.

More generally, the special-shaped signaling between different cell colonys has defined the tissue microenvironment change in tumour, heart and the liver.So, the microscale organization component for research between different cell types, medicine usefulness communication network and be used for standardized, repeatably mode to carry out basic physiological research be important.Yet it is difficult that the tissue that accurately designs a model on microscale is proved to be.

Have and be used for carrying out several technology that heterocyst is cultivated by colony's control.The Transwell plate is in the paracine signaling that is used for traditionally studying between two kinds of different cell colonys.Be used for relying on photoetching technique in the new technology of external imitation tissue microenvironment.A known strategy is based on photoresist or the mask that uses patterning, to allow cell attachment, so that select the zone on surface.Subsequently photoresist or mask are removed the zone that exposes the adhesion of being convenient to second cell type.Second strategy utilizes dielectrophoresis, patterning and cervical cancer cell and white blood cell and red blood cell separated on microelectrode array.Other strategy comprises: cell suspending liquid is directed to the microchannel that different positions is gone up on the surface; Second kind of cell type of permission plantation is to the electricity activation mask that is activated by electricity with the surf zone that allows to adhere to; Organization component with the gravity reinforcement.It is big that these technology prove labor intensive, lacks the control of accurate colony with slowly.Technology based on dielectrophoresis is interesting, because its representative is used for the strategy of cytological classification and microscale tissue reconstruction; Yet its lacks for the spatial control of carrying out the needed accurate cell colony control of quantitative examination, being provided by micro-patterning technology, and depends on and have different polarization characteristics so that collected effectively and the cell of patterning.This has hindered the assurance of uniform cell colony, and wherein this uniform cell colony only can be guaranteed by the molecule selectivity.

In view of above-mentioned content, according to an embodiment of present disclosure assembling, shown in Figure 41-43 at two dimensional tissues.In an exemplary, consider capillary endothelial cell, wherein cell is assembled by becoming PECAM coated with antibody and magnetic bead is suspended in the solution with the endothelial cell that is dissociated on magnetic bead.As shown in figure 41.By using the micro-coil array 202B of IC chip 102, the cell that is attached on the magnetic bead can separatedly be opened, and is directed to then to coated with the structure on the chip surface of fiber adhesion albumen (FN).Especially, shown in Figure 41 (a), two-dimentional endothelial layer is accurately assembled by using micro-coil array 200B, and wherein the fiber adhesion albumen (FN) of micro-patterning is represented with thick black line.In Figure 41 (b), endothelial cell occupies the zone of being represented by little coil of dark color, this slightly coil have the non-zero DC electric current, produce magnetic field thus.In case in place, just allow cell to be adhered to and spread to chip surface, form the individual layer that converges of the geometry of definition.

Subsequently, this endothelial tissue is assembled into " tissue of embedding " in the heart muscle tissue that is pre-existing in.In one embodiment, two-dimentional heart tissue can be fabricated by cultivate newborn mouse ventricular muscle cell on the fiber adhesion albumen of micro-patterning.The cardiac muscle cell of disassociation cultivates in the FN of micro-patterning line, shown in Figure 42 (a).The cardiac muscle cell adheres on the FN line and aims at the FN line, self is assembled into individual layer that converge, anisotropic, and this individual layer can conduct the action potential wavefront.Figure 42 (b) illustrates heart tissue and makes up, and simulates parallel (top) and vertical (bottom) in the kapillary of cardiac fibers.Figure 42 (c) illustrates the interval on the FN island of adhesion plaque (focal-adhesion) size.

By using micro-coil array, capillary endothelial cell can be orientated about the fiber of designed heart tissue and be embedded in the accurate structure, as shown in figure 43.In Figure 43 (a), be guided position between the former cardiac muscle cells by using micro-coil array by the capillary endothelial cell (seeing Figure 43 (b)) of marked by magnetic bead.When locating in position, they keep long enough so that whole being attached on the FN of micro-patterning.As Figure 43 (c) with (d), the bonding FN of endothelial cell, and podocytic process (Lamellipodia) extended to be attached to other island and to adhere to the edge of cardiac muscle cell's FN line on it.

The FN island of adhesion plaque size may be unfavorable for that the myocyte adheres to and spreads, and tears them before they can fully adhere to because these myocytes' spontaneous contraction makes from single little FN island.Yet, bonding these islands of capillary endothelial cell and with podocytic process extend to be extended to occupy simultaneously several.Therefore, there is the zone on little FN island can optionally handle endothelial cell rather than cardiac muscle cell's (seeing Figure 42) by micro-patterning.The endothelial cell that is attached on the magnetic bead adds one at a time by micro-coil array, as shown in figure 43, because after the myocyte is attached on the substrate they are placed in the solution, can cause along mixing, the unsteered colony of the FN line of micro-patterning.In the cultivation of 24 hours or shorter time, allow endothelium embedding thing (embeds) diffusion that makes up.Immunohistochemistry can be used for labeled cell, to follow the tracks of when making up the back at specific time point based on micro-coil array their growth.Especially, tissue can (sarcomeric α-actinin), PECAM and nuclear DNA (DAPI) be by triple staining at the band αFu Jidongdanbai, so that accurately be positioned at the separatrix between endothelial cell and the cardiac muscle cell, and check endothelial cell possible migrates and breeds.On the one hand, the media condition of improvement makes endothelial cell proliferation minimize, but supports endothelial cell expansion and myocyte to beat.

According to above-mentioned methodology, can finish homogeneity and geometry degree of accuracy that endothelial cell embeds, and the invasion that stops endothelial cell between the cardiac muscle fiber.The applicant recognizes and understands, and the cardiac muscle cell being placed on the surface of micro-patterning before the assembling endothelium embeds is important step in the processing procedure.Especially, compared with other cell type, the cardiac muscle cell needs more time to be attached to and is adapted to the extracellular matrix embolus.In addition, capillary endothelial cell is quite migration, and the cardiac muscle cell is not.Therefore, by placing the cardiac muscle cell in advance, the cell that endothelium embeds can be pinned in after assembling in the zone of their appointments effectively.

Conclusion

The various embodiment of commingled system discussed here merge electromagnetics, miniflow, semiconductor physics, lithography technique, high frequency (for example, RF) FEEDBACK CONTROL and the biology in electronics, analog/digital integrated circuit, the complementary system.In various exemplary, such commingled system can be configured to " biochip ", the general programmable device that can carry out various Bioexperiment on sub-micron scale is provided, helps realizing industry, science and military exploitation of interested " laboratory on the sheet " thus greatly.

After describing several illustrative embodiment in this wise, should be appreciated that those skilled in the art make various changes, modification and improvement easily.Such change, modification and improvement are intended to the part as present disclosure, and are intended to belong in the spirit and scope of present disclosure.Though some example given here relates to the specific combination of function or structural unit, should be appreciated that these functions and unit can otherwise be combined to finish identical and different purposes according to the present invention.Especially, do not plan to be excluded in other embodiments similarly in conjunction with step, unit and the characteristic of an embodiment discussion or beyond other the role.Therefore, above explanation and accompanying drawing do not limit and be not intended to conduct as just example.

Claims

1. equipment comprises:

The field that a plurality of CMOS make generates parts;

Micro-fluidic system, it is configured to be included near the fluid of field generation parts that a plurality of CMOS make; And

At least one controller, it is configured to control the field generation parts that a plurality of CMOS make, to generate at least one electric field with enough intensity or magnetic field, so that interact with at least one sample that is suspended in the fluid.

2. according to the equipment of claim 1, wherein at least one controller is configured to control the field generation parts that a plurality of CMOS make, with generate a plurality of have enough intensity, on programmable, the space or the time go up variable electric field or magnetic field so that interact with at least one sample that is suspended in the fluid.

3. according to the equipment of claim 2, further comprise at least one processor that is coupled at least one controller, described at least one processor is configured to control described at least one controller, so that carry out in manipulation, detection, imaging and the sign of described at least one sample at least one via described a plurality of electric fields or magnetic field.

4. according to the equipment of claim 3, wherein said at least one processor is configured to be convenient to carry out based on the detection of described at least one sample the programmable unmanned operation of described at least one sample.

5. according to the equipment of claim 1, wherein said at least one controller comprises the field control assembly that a plurality of CMOS make, and the field that these parts and a plurality of CMOS make generates parts and together forms integrated circuit (IC) chip.

6. according to the equipment of claim 5, wherein said micro-fluidic system integrally is coupled with described integrated circuit (IC) chip, to form CMOS/ miniflow commingled system.

7. according to the equipment of claim 6, wherein micro-fluidic system comprises at least one polyimide layer, and its field that is deposited over described CMOS making generates on the parts, forms at least one microchannel or liquid storage pool in this layer.

8. according to the equipment of claim 6, wherein said micro-fluidic system comprises at least one epoxy resin layer, and its field that is deposited over described CMOS making generates on the parts, forms at least one microchannel or liquid storage pool in this layer.

9. according to the equipment of claim 6, wherein said micro-fluidic system comprises at least one dimethyl silicone polymer (PDMS) mould, and its field that is deposited over described CMOS making generates on the parts, forms at least one microchannel or liquid storage pool in this mould.

10. according to the equipment of claim 5, wherein said a plurality of control assemblies comprise:

A plurality of programmable exchanges or multiple connection parts; And

A plurality of current sources or voltage source.

11. equipment according to claim 10, wherein said a plurality of control assemblies further comprise a plurality of high-frequency detection parts, and carry out at least one in the detection, imaging and the sign that are suspended in described at least one sample in the fluid in its at least one electric field or magnetic field that is configured to be convenient to via described generation.

12. according to the equipment of claim 11, further comprise the temperature controlling unit that at least one CMOS makes, the field that the field control assembly that itself and described a plurality of CMOS make and described a plurality of CMOS make generates parts and together forms described integrated circuit (IC) chip.

13. equipment according to claim 12, further comprise at least one processor that is coupled to described at least one controller, described at least one processor is configured to control described at least one controller, so that carry out in manipulation, detection, imaging and the sign of described at least one sample at least one via at least one electric field that is generated or magnetic field.

14. according to the equipment of claim 13, wherein said at least one processor is configured to be convenient to carry out programmable unmanned operation for described at least one sample based on the detection of described at least one sample.

15. according to the equipment of claim 1, the field that wherein said a plurality of CMOS make generates parts and comprises a plurality of little coils.

16. according to the equipment of claim 15, wherein said a plurality of little coils are aligned to two-dimensional array.

17. according to the equipment of claim 15, wherein each little coil comprise at least two of conductor ring concentric in the axial direction, separated portions spatially.

18. according to the equipment of claim 15, wherein said at least one controller comprises a plurality of current sources or voltage source and a plurality of exchange or the multiple connection parts that are coupled to described a plurality of little coils.

19. according to the equipment of claim 18, wherein said at least one controller further comprises a plurality of radio frequencies (RF) detection part that is coupled to described a plurality of little coils.

20. according to the equipment of claim 19, wherein said a plurality of RF detection parts comprise frequency-locked loop, it is configured to be convenient to, and described at least one sample in the fluid detects for being suspended in, imaging and sign at least one.

21. equipment according to claim 20, wherein said frequency-locked loop comprises at least one bridge diagram, described at least one bridge diagram comprises at least one little coil of described a plurality of little coils, described at least one bridge diagram is configured to and generates at least one signal, and this signal representative is owing near the change of the inductance of described at least one the little coil that exists described at least one sample to cause described at least one little coil.

22. a method may further comprise the steps:

A) generate parts from the field that a plurality of CMOS make and generate at least one electric field or magnetic field, described at least one electric field or magnetic field, has enough intensity, so that interact with at least one sample that is suspended in the fluid, the field that wherein said fluid is comprised in a plurality of CMOS making generates near the micro-fluidic system of parts.

23. according to the method for claim 22, wherein said steps A) may further comprise the steps:

A1) generate a plurality of have sufficient intensity programmable, spatially or the time go up variable electric field or magnetic field so that interact with described at least one sample that is suspended in the described fluid.

24. the method according to claim 23 further may further comprise the steps:

B) control described a plurality of electric fields or magnetic field, so as for described at least one sample handle, in detection, imaging and the sign at least one.

25. according to the method for claim 24, wherein said step B) may further comprise the steps:

Control described a plurality of electric field or magnetic field, so that carry out unmanned operation for described at least one sample based on the detection of described at least one sample.

26. according to the method for claim 24, wherein said steps A 1) may further comprise the steps:

Via a plurality of programmable exchanges or multiple connection parts voltage or electric current are applied on the field generation parts of described a plurality of CMOS making.

27. according to the method for claim 24, wherein said steps A 1) may further comprise the steps:

A2) apply at least one high-frequency signal and generate at least one generation parts of parts to the field that described a plurality of CMOS make, so as at least one sample detect, in imaging and the sign at least one.

28. according to the method for claim 27, wherein said steps A 2) may further comprise the steps:

Monitor the frequency of described at least one high-frequency signal, there is or does not exist described at least one sample in wherein said frequency indication near described at least one generation parts.

29., further comprising the steps of according to the method for claim 27:

C) temperature of described at least one sample of adjustment.

30. according to the method for claim 22, the field that wherein said a plurality of CMOS make generates parts and comprises a plurality of little coils, each little coil comprise at least two of conductor ring concentric in the axial direction, separated portions spatially.